Quinazoline Derivatives as VEGF Inhibitors

ABSTRACT

The invention relates to quinazoline derivatives of formula (I), wherein m is an integer from 1 to 3; R 1  represents halogeno or C 1-3 alkyl; X 1  represents —O—; R 2  is selected from one of the following three groups: 1) C 1-5 alkylR 3  (wherein R 3  is piperidin-4-yl which may bear one or two substituents selected from hydroxy, halogeno, C 1-4 alkyl, C 1-4 hydroxyalkyl and C 1-4 alkoxy; 2) C 2-5 alkenylR 3  (wherein R 3  is as defined hereinbefore); 3) C 2-5 alkynylR 3  (wherein R 3  is as defined hereinbefore); and wherein any alkyl, alkenyl or alkynyl group may bear one or more substituents selected from hydroxy, halogeno and amino; and salts thereof; processes for their preparation, pharmaceutical compositions containing a compound of formula (I) or a pharmaceutically acceptable salt thereof as active ingredient. The compounds of formula (I) and the pharmaceutically acceptable salts thereof inhibit the effects of VEGF, a property of value in the treatment of a number of disease states including cancer and rheumatoid arthritis.

The present invention relates to quinazoline derivatives, processes fortheir preparation, pharmaceutical compositions containing them as activeingredient, methods for the treatment of disease states associated withangiogenesis and/or increased vascular permeability, to their use asmedicaments and to their use in the manufacture of medicaments for usein the production of antiangiogenic and/or vascular permeabilityreducing effects in warm-blooded animals such as humans.

Normal angiogenesis plays an important role in a variety of processesincluding embryonic development, wound healing and several components offemale reproductive function. Undesirable or pathological angiogenesishas been associated with disease states including diabetic retinopathy,psoriasis, cancer, rheumatoid arthritis, atheroma, Kaposi's sarcoma andhaemangioma (Fan et al, 1995, Trends Pharmacol. Sci. 16: 57-66; Folkman,1995, Nature Medicine 1: 27-31). Alteration of vascular permeability isthought to play a role in both normal and pathological physiologicalprocesses (Cullinan-Bove et al, 1993, Endocrinology 133: 829-837; Sengeret al, 1993, Cancer and Metastasis Reviews, 12: 303-324). Severalpolypeptides with in vitro endothelial cell growth promoting activityhave been identified including, acidic and basic fibroblast growthfactors (aFGF & bFGF) and vascular endothelial growth factor (VEGF). Byvirtue of the restricted expression of its receptors, the growth factoractivity of VEGF, in contrast to that of the FGFs, is relativelyspecific towards endothelial cells. Recent evidence indicates that VEGFis an important stimulator of both normal and pathological angiogenesis(Jakeman et al, 1993, Endocrinology, 133: 848-859; Kolch et al, 1995,Breast Cancer Research and Treatment, 36:139-155) and vascularpermeability (Connolly et al, 1989, J. Biol. Chem. 264: 20017-20024).Antagonism of VEGF action by sequestration of VEGF with antibody canresult in inhibition of tumour growth (Kim et al, 1993, Nature 362:841-844).

Receptor tyrosine kinases (RTKs) are important in the transmission ofbiochemical signals across the plasma membrane of cells. Thesetransmembrane molecules characteristically consist of an extracellularligand-binding domain connected through a segment in the plasma membraneto an intracellular tyrosine kinase domain. Binding of ligand to thereceptor results in stimulation of the receptor-associated tyrosinekinase activity which leads to phosphorylation of tyrosine residues onboth the receptor and other intracellular molecules. These changes intyrosine phosphorylation initiate a signalling cascade leading to avariety of cellular responses. To date, at least nineteen distinct RTKsubfamilies, defined by amino acid sequence homology, have beenidentified. One of these subfamilies is presently comprised by thefms-like tyrosine kinase receptor, Flt or Fltl, the kinase insertdomain-containing receptor, KDR (also referred to as Flk-1), and anotherfms-like tyrosine kinase receptor, Flt4. Two of these related RTKs, Fltand KDR, have been shown to bind VEGF with high affinity (De Vries etal, 1992, Science 255: 989-991; Terman et al, 1992, Biochem. Biophys.Res. Comm. 1992, 187: 1579-1586). Binding of VEGF to these receptorsexpressed in heterologous cells has been associated with changes in thetyrosine phosphorylation status of cellular proteins and calcium fluxes.

Quinazoline derivatives which are inhibitors of VEGF receptor tyrosinekinase are described in International Patent Applications PublicationNos. WO 97/30035 and WO 98/13354. In WO 97/30035 and WO 98/13354compounds are described which possess activity against VEGF receptortyrosine kinase whilst possessing some activity against EGF receptortyrosine kinase.

Compounds of the present invention fall within the broad generaldisclosure of WO 97/30035 and WO 98/13354. We have found that compoundsof the present invention possess very good inhibitory activity againstVEGF receptor tyrosine kinase. Compounds of the present invention, whichhave been tested, show in vivo activity against a range of tumourxenografts in mice. Compounds of the present invention possess abeneficial toxicological profile when tested over 14 days in rats.Compounds of the present invention possess very good inhibitory activityagainst VEGF receptor tyrosine kinase, show in vivo activity against arange of tumour xenografts in mice and possess a beneficialtoxicological profile when tested over 14 days in rats.

Compounds of the present invention inhibit the effects of VEGF, aproperty of value in the treatment of disease states associated withangiogenesis and/or increased vascular permeability such as cancer,diabetes, psoriasis, rheumatoid arthritis, Kaposi's sarcoma,haemangioma, acute and chronic nephropathies, atheroma, arterialrestenosis, autoimmune diseases, acute inflammation, excessive scarformation and adhesions, endometriosis, dysfunctional uterine bleedingand ocular diseases with retinal vessel proliferation.

Compounds of the present invention possess good activity against VEGFreceptor tyrosine kinase whilst possessing some activity against EGFreceptor tyrosine kinase. Furthermore, some compounds of the presentinvention, possess substantially higher potency against VEGF receptortyrosine kinase than against EGF receptor tyrosine kinase or FGF R1receptor tyrosine kinase. While we do not wish to be bound bytheoretical considerations such compounds may for example be of interestin treating tumours which are associated with VEGF, especially thosetumours which are dependent on VEGF for their growth. It is furtherbelieved that these compounds may be of interest in treating tumourstates associated with both VEGF and EGF, especially where a patient issuffering from a condition in which tumours are present which aredependent on both VEGF and EGF for their growth.

According to one aspect of the present invention there is provided aquinazoline derivative of the formula I:

-   -   wherein:

-   m is an integer from 1 to 3;

-   R¹ represents halogeno or C₁₋₃alkyl;

-   X¹ represents —O—;

-   R² is selected from one of the following three groups:

-   1) C,_(—5)alkylR³ (wherein R³ is piperidin-4-yl which may bear one    or two substituents selected from hydroxy, halogeno, C₁₋₄alkyl,    C₁₋₄hydroxyalkyl and C₁₋₄alkoxy;

-   2) C₂₋₅alkenylR³ (wherein R³ is as defined hereinbefore);

-   3) C₂₋₅alkynylR³ (wherein R³ is as defined hereinbefore); and    wherein any alkyl, alkenyl or alkynyl group may bear one or more    substituents selected from hydroxy, halogeno and amino;

-   or a salt thereof or a prodrug thereof.

-   Preferably m is 2.

-   Preferably the phenyl group bearing (R¹)_(m) is selected from    2-fluoro-4-methylphenyl, 4-chloro-2,6-difluorophenyl,    4-bromo-2,6-difluorophenyl, 4-chloro-2-fluorophenyl group and    4-bromo-2-fluorophenyl.

-   More preferably the phenyl group bearing (R¹)_(m) is selected from    4-chloro-2-fluorophenyl and 4-bromo-2-fluorophenyl.

-   Most preferably the phenyl group bearing (R¹)_(m) is    4-bromo-2-fluorophenyl. Preferably R² is C₁₋₅alkylR³ (wherein R³ is    as defined hereinbefore).

-   More preferably R² is C₁₋₃alkylR³(wherein R³ is as defined    hereinbefore). Particularly R² is piperidin-4-ylmethyl in which the    piperidine ring may bear one or two substituents as defined    hereinbefore.

-   More particularly R² is piperidin-4-ylmethyl in which the piperidine    ring may bear one or two substituents selected from C₁₋₄alkyl.

-   Especially R² is 1-methylpiperidin-4-ylmethyl.

According to a further aspect of the present invention there is provideda quinazoline derivative of the formula II:

wherein:

-   ma is an integer from 1 to 3;-   R^(1a) represents halogeno or C₁₋₃alkyl;-   X^(1a) represents —O—;-   R^(2a) is selected from one of the following three groups:-   1) C₁₋₅alkylR³ (wherein R³ is as defined hereinbefore);-   2) C₂₋₅alkenylR³ (wherein R³ is as defined hereinbefore);-   3) C₂₋₅alkynylR³ (wherein R³ is as defined hereinbefore);-   or a salt thereof or a prodrug thereof.-   Preferably ma is 2.-   Preferably the phenyl group bearing (R^(1a))_(ma) is selected from    2-fluoro-4-methylphenyl, 4-chloro-2,6-difluorophenyl,    4-bromo-2,6-difluorophenyl, 4-chloro-2-fluorophenyl group and    4-bromo-2-fluorophenyl.-   More preferably the phenyl group bearing (R^(1a))_(ma) is selected    from 4-chloro-2-fluorophenyl and 4-bromo-2-fluorophenyl.-   Most preferably the phenyl group bearing (R^(1a))_(ma) is    4-bromo-2-fluorophenyl.-   Preferably R^(2a) is C₁₋₅alkylR³ (wherein R³ is as defined    hereinbefore).-   More preferably R^(2a) is C₁₋₃alkylR³(wherein R³ is as defined    hereinbefore).-   Particularly le is piperidin-4-ylmethyl in which the piperidine ring    may bear one or two substituents as defined hereinbefore.-   More particularly R^(2a) is piperidin-4-ylmethyl in which the    piperidine ring may bear one or two substituents selected from    C₁₋₄alkyl.-   Especially R^(2a) is 1-methylpiperidin-4-ylmethyl.-   Preferred compounds of the present invention include:-   4-(4-chloro-2-fluoroanilino)-6-methoxy-7-(1    -methylpiperidin-4-ylmethoxy)quinazoline,-   4-(2-fluoro-4-methylanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline,-   4-(4-bromo-2-fluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline,-   4-(4-chloro-2,6-difluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline,-   4-(4-bromo-2,6-difluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline,-   4-(4-chloro-2-fluoroanilino)-6-methoxy-7-(piperidin-4-ylmethoxy)quinazoline,-   4-(2-fluoro-4-methylanilino)-6-methoxy-7-(piperidin-4-ylmethoxy)quinazoline,-   4-(4-bromo-2-fluoroanilino)-6-methoxy-7-(piperidin-4-ylmethoxy)quinazoline,-   4-(4-chloro-2,6-difluoroanilino)-6-methoxy-7-(piperidin-4-ylmethoxy)quinazoline,    and-   4-(4-bromo-2,6-difluoroanilino)-6-methoxy-7-(piperidin-4-ylmethoxy)quinazoline,-   and salts thereof especially hydrochloride salts thereof.-   More preferred compounds of the present invention include:-   4-(4-chloro-2-fluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline,-   4-(4-bromo-2-fluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline,-   4-(4-chloro-2,6-difluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline,-   4-(4-bromo-2,6-difluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline,-   4-(4-chloro-2-fluoroanilino)-6-methoxy-7-(piperidin-4-ylmethoxy)quinazoline,-   4-(4-bromo-2-fluoroanilino)-6-methoxy-7-(piperidin-4-ylmethoxy)quinazoline,-   4-(4-chloro-2,6-difluoroanilino)-6-methoxy-7-(piperidin-4-ylmethoxy)quinazoline,    and-   4-(4-bromo-2,6-difluoroanilino)-6-methoxy-7-(piperidin-4-ylmethoxy)quinazoline,    and salts thereof especially hydrochloride salts thereof.-   Particularly preferred compounds of the present invention include:-   4-(4-chloro-2-fluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline,-   4-(4-bromo-2-fluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline,-   4-(4-chloro-2,6-difluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline,    and-   4-(4-bromo-2,6-difluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline,-   and salts thereof especially hydrochloride salts thereof.-   More particularly preferred compounds of the present invention    include:-   4-(4-chloro-2-fluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline    and-   4-(4-bromo-2-fluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline,-   and salts thereof especially hydrochloride salts thereof.-   An especially preferred compound of the present invention is    4-(4-bromo-2-fluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline    and salts thereof especially hydrochloride salts thereof.

For the avoidance of doubt it is to be understood that where in thisspecification a group is qualified by ‘hereinbefore defined’ or ‘definedhereinbefore’ the said group encompasses the first occurring andbroadest definition as well as each and all of the preferred definitionsfor that group. And a similar convention applies to ‘hereinafterdefined’ or ‘defined hereinafter’.

In this specification unless stated otherwise the term “alkyl” includesboth straight and branched chain alkyl groups but references toindividual alkyl groups such as “propyl” are specific for the straightchain version only. An analogous convention applies to other genericterms. Unless otherwise stated the term “alkyl” advantageously refers tochains with 1-5 carbon atoms, preferably 1-3 carbon atoms. The term“alkoxy” as used herein, unless stated otherwise includes “alkyl”-O—groups in which “alkyl” is as hereinbefore defined. The term “aryl” asused herein unless stated otherwise includes reference to a C₆₋₁₀ arylgroup which may, if desired, carry one or more substituents selectedfrom halogen, alkyl, alkoxy, nitro, trifluoromethyl and cyano, (whereinalkyl and alkoxy are as hereinbefore defined). The term “aryloxy” asused herein unless otherwise stated includes “aryl” —O-groups in which“aryl” is as hereinbefore defined. The term “sulphonyloxy” as usedherein refers to alkylsulphonyloxy and arylsulphonyloxy groups in which“alkyl” and “aryl” are as hereinbefore defined. The term “alkanoyl” asused herein unless otherwise stated includes formyl and alkylC═O groupsin which “alkyl” is as defined hereinbefore, for example C₂alkanoyl isethanoyl and refers to CH₃C═O, C₁alkanoyl is formyl and refers to CHO.In this specification unless stated otherwise the term “alkenyl”includes both straight and branched chain alkenyl groups but referencesto individual alkenyl groups such as 2-butenyl are specific for thestraight chain version only. Unless otherwise stated the term “alkenyl”advantageously refers to chains with 2-5 carbon atoms, preferably 3-5carbon atoms. In this specification unless stated otherwise the term“alkynyl” includes both straight and branched chain alkynyl groups butreferences to individual alkynyl groups such as 2-butynyl are specificfor the straight chain version only. Unless otherwise stated the term“alkynyl” advantageously refers to chains with 2-5 carbon atoms,preferably 3-5 carbon atoms.

In formula I, as hereinbefore defined, hydrogen will be present atpositions 2, 5 and 8 of the quinazoline group.

Within the present invention it is to be understood that a compound ofthe formula I or a salt thereof may exhibit the phenomenon oftautomerism and that the formulae drawings within this specification canrepresent only one of the possible tautomeric forms. It is to beunderstood that the invention encompasses any tautomeric form whichinhibits VEGF receptor tyrosine kinase activity and is not to be limitedmerely to any one tautomeric form utilised within the formulae drawings.

It is also to be understood that certain compounds of the formula I andsalts thereof can exist in solvated as well as unsolvated forms such as,for example, hydrated forms. It is to be understood that the inventionencompasses all such solvated forms which inhibit VEGF receptor tyrosinekinase activity.

For the avoidance of any doubt, it is to be understood that in acompound of the formula I when R² is, for example, a group of formulaC₂₋₅alkenylR³ it is the C₂₋₅alkenyl moiety which is bound to X¹ and ananalogous convention applies to other groups. When R² is a group1-R³prop-1-en-3-yl it is the first carbon to which the group R³ isattached and it is the third carbon which is linked to X¹, similarlywhen R² is a group 2-R³pent-3-en-5-yl it is the second carbon to whichthe group R³ is attached and it is the fifth carbon which is linked toX¹, and an analogous convention applies to other groups.

Compounds of Formula I may be administered in the form of a prodrugwhich is broken down in the human or animal body to give a compound ofthe Formula I. Examples of prodrugs include in vivo hydrolysable estersof a compound of the Formula I.

Various forms of prodrugs are known in the art. For examples of suchprodrug derivatives see:

-   a) Design of Prodrugs, edited by H. Bundgaard, (Elsevier, 1985) and    Methods in Enzymology, Vol. 42, p. 309-396, edited by K. Widder, et    al. (Academic Press, 1985);-   b) A Textbook of Drug Design and Development, edited by    Krogsgaard-Larsen and H. Bundgaard, Chapter 5 “Design and    Application of Prodrugs”, by H. Bundgaard p. 113-191 (1991);-   c) H. Bundgaard, Advanced Drug Delivery Reviews, 8, 1-38 (1992);-   d) H. Bundgaard, et al., Journal of Pharmaceutical Sciences, 77, 285    (1988); and-   e) N. Kakeya, et al., Chem Pharm Bull, 32, 692 (1984).

An in vivo hydrolysable ester of a compound of Formula I containing ahydroxy group includes inorganic esters such as phosphate esters(including phosphoramidic cyclic esters) and a-acyloxyalkyl ethers andrelated compounds which as a result of the in vivo hydrolysis of theester breakdown to give the parent hydroxy group/s. Examples ofa-acyloxyalkyl ethers include acetoxymethoxy and2,2-dimethylpropionyloxy-methoxy. A selection of in vivo hydrolysableester forming groups for hydroxy include alkanoyl, benzoyl, phenylacetyland substituted benzoyl and phenylacetyl, alkoxycarbonyl (to give alkylcarbonate esters), dialkylcarbamoyl andN-(dialkylaminoethyl)-N-alkylcarbamoyl (to give carbamates),dialkylaminoacetyl and carboxyacetyl. Examples of substituents onbenzoyl include morpholino and piperazino linked from a ring nitrogenatom via a methylene group to the 3- or 4- position of the benzoyl ring.

The present invention relates to the compounds of formula I ashereinbefore defined as well as to the salts thereof. Salts for use inpharmaceutical compositions will be pharmaceutically acceptable salts,but other salts may be useful in the production of the compounds offormula I and their pharmaceutically acceptable salts. Pharmaceuticallyacceptable salts of the invention may, for example, include acidaddition salts of the compounds of formula I as hereinbefore definedwhich are sufficiently basic to form such salts. Such acid additionsalts include for example salts with inorganic or organic acidsaffording pharmaceutically acceptable anions such as with hydrogenhalides (especially hydrochloric or hydrobromic acid of whichhydrochloric acid is particularly preferred) or with sulphuric orphosphoric acid, or with trifluoroacetic, citric or maleic acid. Inaddition where the compounds of formula I are sufficiently acidic,pharmaceutically acceptable salts may be formed with an inorganic ororganic base which affords a pharmaceutically acceptable cation. Suchsalts with inorganic or organic bases include for example an alkalimetal salt, such as a sodium or potassium salt, an alkaline earth metalsalt such as a calcium or magnesium salt, an ammonium salt or forexample a salt with methylamine, dimethylamine, trimethylamine,piperidine, morpholine or tris-(2-hydroxyethyl)amine.

A compound of the formula I, or salt thereof, and other compounds of theinvention (as hereinafter defined) may be prepared by any process knownto be applicable to the preparation of chemically-related compounds.Such processes include, for example, those illustrated in EuropeanPatent Applications, Publication Nos. 0520722, 0566226, 0602851 and0635498 and in International Patent Applications Publication Nos. WO97/22596, WO 97/30035, WO 97/32856 and WO 98/13354. Such processes, areprovided as a further feature of the invention and are as describedhereinafter. Necessary starting materials may be obtained by standardprocedures of organic chemistry. The preparation of such startingmaterials is described within the accompanying non-limiting Examples.Alternatively necessary starting materials are obtainable by analogousprocedures to those illustrated which are within the ordinary skill ofan organic chemist.

Thus the following processes (a) to (d) and (i) to (iv) constitutefurther features of the present invention.

Synthesis of Compounds of Formula I

(a) Compounds of the formula I and salts thereof may be prepared by thereaction of a compound of the formula III:

(wherein R² and X¹ are as defined hereinbefore and L¹ is a displaceablemoiety), with a compound of the formula IV:

(wherein R¹ and m are as defined hereinbefore) whereby to obtaincompounds of the formula I and salts thereof. A convenient displaceablemoiety L¹ is, for example, a halogeno, alkoxy (preferably C₁₋₄alkoxy),aryloxy or sulphonyloxy group, for example a chloro, bromo, methoxy,phenoxy, methanesulphonyloxy or toluene-4-sulphonyloxy group.

The reaction is advantageously effected in the presence of either anacid or a base. Such an acid is, for example, an anhydrous inorganicacid such as hydrogen chloride. Such a base is, for example, an organicamine base such as, for example, pyridine, 2,6-lutidine, collidine,4-dimethylaminopyridine, triethylamine, morpholine, N-methylmorpholineor diazabicyclo[5.4.0]undec-7-ene, or for example, an alkali metal oralkaline earth metal carbonate or hydroxide, for example sodiumcarbonate, potassium carbonate, calcium carbonate, sodium hydroxide orpotassium hydroxide. Alternatively such a base is, for example, analkali metal hydride, for example sodium hydride, or an alkali metal oralkaline earth metal amide, for example sodium amide or sodiumbis(trimethylsilyl)amide. The reaction is preferably effected in thepresence of an inert solvent or diluent, for example an alkanol or estersuch as methanol, ethanol, 2-propanol or ethyl acetate, a halogenatedsolvent such as methylene chloride, trichloromethane or carbontetrachloride, an ether such as tetrahydrofuran or 1,4-dioxan, anaromatic hydrocarbon solvent such as toluene, or a dipolar aproticsolvent such as N,N-dimethylformamide, N,N-dimethylacetamide,N-methylpyrrolidin-2-one or dimethylsulphoxide. The reaction isconveniently effected at a temperature in the range, for example, 10 to150° C., preferably in the range 20 to 80° C.

The compound of the invention may be obtained from this process in theform of the free base or alternatively it may be obtained in the form ofa salt with the acid of the formula H-L¹ wherein L¹ has the meaningdefined hereinbefore. When it is desired to obtain the free base fromthe salt, the salt may be treated with a base as defined hereinbeforeusing a conventional procedure.

(b) Compounds of the formula I and salts thereof may be prepared by thereaction, conveniently in the presence of a base as definedhereinbefore, of a compound of the formula V:

(wherein m, X¹ and R¹ are as hereinbefore defined) with a compound offormula VI:

R²-L¹   (VI)

(wherein R² and L¹ are as hereinbefore defined); L¹ is a displaceablemoiety for example a halogeno or sulphonyloxy group such as a bromo ormethanesulphonyloxy group. Conveniently L¹ is a group O—⁺P(Y)₃ (whereinY is butyl or phenyl) and in such cases the compound of formula VI isconveniently formed in situ. The reaction is preferably effected in thepresence of a base (as defined hereinbefore in process (a)) andadvantageously in the presence of an inert solvent or diluent (asdefined hereinbefore in process (a)), advantageously at a temperature inthe range, for example 10 to 150° C., conveniently at about 50° C.

(c) Compounds of the formula I and salts thereof may be prepared by thereaction of a compound of the formula VII:

with a compound of the formula VIII:

R²—X¹—H   (VIII)

(wherein L¹, R¹, R², m and X¹ are all as hereinbefore defined). Thereaction may conveniently be effected in the presence of a base (asdefined hereinbefore in process (a)) and advantageously in the presenceof an inert solvent or diluent (as defined hereinbefore in process (a)),advantageously at a temperature in the range, for example 10 to 150° C.,conveniently at about 100° C.

(d) Compounds of the formula I and salts thereof may be prepared by thedeprotection of a compound of the formula IX:

wherein R¹, m and X¹ are all as hereinbefore defined, and R⁴ representsa protected R² group wherein R² is as defined hereinbefore butadditionally bears one or more protecting groups P². The choice ofprotecting group P² is within the standard knowledge of an organicchemist, for example those included in standard texts such as“Protective Groups in Organic Synthesis” T. W. Greene and R. G. M. Wuts,2nd Ed. Wiley 1991. Preferably P² is a protecting group such as acarbamate (alkoxycarbonyl) (such as, for example, tert-butoxycarbonyl,tert-amyloxycarbonyl, cyclobutoxycarbonyl, propoxycarbonyl,methoxycarbonyl, ethoxycarbonyl, isopropoxycarbonyl, allyloxycarbonyl orbenzyloxycarbonyl). More preferably P² is tert-butoxycarbonyl. Thereaction is preferably effected in the presence of an acid. Such an acidis, for example, an inorganic acid such as hydrogen chloride, hydrogenbromide or an organic acid such as trifluoroacetic acid,trifluoromethane sulphonic acid. The reaction may be effected in thepresence of an inert solvent such as methylene chloride,trichloromethane and in the presence of a trace of water. The reactionis conveniently effected at a temperature in the range, for example,10-100° C., preferably in the range 20-80° C.

Synthesis of Intermediates

(i) The compounds of formula III and salts thereof in which L¹ ishalogeno may for example be prepared by halogenating a compound of theformula X:

(wherein R² and X¹ are as hereinbefore defined).

Convenient halogenating agents include inorganic acid halides, forexample thionyl chloride, phosphorus(III)chloride,phosphorus(V)oxychloride and phosphorus(V)chloride. The halogenationreaction is conveniently effected in the presence of an inert solvent ordiluent such as for example a halogenated solvent such as methylenechloride, trichloromethane or carbon tetrachloride, or an aromatichydrocarbon solvent such as benzene or toluene. The reaction isconveniently effected at a temperature in the range, for example 10 to150° C., preferably in the range 40 to 100° C.

The compounds of formula X and salts thereof may for example be preparedby reacting a compound of the formula XI:

(wherein L¹ is as hereinbefore defined) with a compound of the formulaVIII as hereinbefore defined. The reaction may conveniently be effectedin the presence of a base (as defined hereinbefore in process (a)) andadvantageously in the presence of an inert solvent or diluent (asdefined hereinbefore in process (a)), advantageously at a temperature inthe range, for example 10 to 150° C., conveniently at about 100° C.

The compounds of formula X and salts thereof may also be prepared bycyclising a compound of the formula XII:

(wherein R² and X¹, are as hereinbefore defined, and A¹ is an hydroxy,alkoxy (preferably alkoxy) or amino group) whereby to form a compound offormula X or salt thereof. The cyclisation may be effected by reacting acompound of the formula XII, where A¹ is an hydroxy or alkoxy group,with formamide or an equivalent thereof effective to cause cyclisationwhereby a compound of formula X or salt thereof is obtained, such as[3-(dimethylamino)-2-azaprop-2-enylidene]dimethylammonium chloride. Thecyclisation is conveniently effected in the presence of formamide assolvent or in the presence of an inert solvent or diluent such as anether for example 1,4-dioxan. The cyclisation is conveniently effectedat an elevated temperature, preferably in the range 80 to 200° C. Thecompounds of formula X may also be prepared by cyclising a compound ofthe formula XII, where A′ is an amino group, with formic acid or anequivalent thereof effective to cause cyclisation whereby a compound offormula X or salt thereof is obtained. Equivalents of formic acideffective to cause cyclisation include for example atri-C₁₋₄alkoxymethane, for example triethoxymethane andtrimethoxymethane. The cyclisation is conveniently effected in thepresence of a catalytic amount of an anhydrous acid, such as a sulphonicacid for example p-toluenesulphonic acid, and in the presence of aninert solvent or diluent such as for example a halogenated solvent suchas methylene chloride, trichloromethane or carbon tetrachloride, anether such as diethyl ether or tetrahydrofuran, or an aromatichydrocarbon solvent such as toluene. The cyclisation is convenientlyeffected at a temperature in the range, for example 10 to 100° C.,preferably in the range 20 to 50° C.

Compounds of formula XII and salts thereof may for example be preparedby the reduction of the nitro group in a compound of the formula XIII:

(wherein R², X¹ and A¹ are as hereinbefore defined) to yield a compoundof formula XII as hereinbefore defined. The reduction of the nitro groupmay conveniently be effected by any of the procedures known for such atransformation. The reduction may be carried out, for example, by thehydrogenation of a solution of the nitro compound in the presence of aninert solvent or diluent as defined hereinbefore in the presence of ametal effective to catalyse hydrogenation reactions such as palladium orplatinum. A further reducing agent is, for example, an activated metalsuch as activated iron (produced for example by washing iron powder witha dilute solution of an acid such as hydrochloric acid). Thus, forexample, the reduction may be effected by heating the nitro compound andthe activated metal in the presence of a solvent or diluent such as amixture of water and alcohol, for example methanol or ethanol, to atemperature in the range, for example 50 to 150° C., conveniently atabout 70° C.

Compounds of the formula XIII and salts thereof may for example beprepared by the reaction of a compound of the formula XIV:

(wherein L¹ and A¹ are as hereinbefore defined) with a compound of theformula VIII as hereinbefore defined to give a compound of the formualXIII. The reaction of the compounds of formulae XIV and VIII isconveniently effected under conditions as described for process (c)hereinbefore.

Compounds of formula XIII and salts thereof, may for example also beprepared by the reaction of a compound of the formula XV:

(wherein X¹ and A¹ are as hereinbefore defined) with a compound of theformula VI as hereinbefore defined to yield a compound of formula XIIIas hereinbefore defined. The reaction of the compounds of formulae XVand VI is conveniently effected under conditions as described forprocess (b) hereinbefore.

The compounds of formula III and salts thereof may also be prepared forexample by reacting a compound of the formula XVI:

(wherein X¹ is as hereinbefore defined and L² represents a displaceableprotecting moiety) with a compound of the formula VI as hereinbeforedefined, whereby to obtain a compound of formula III in which L¹ isrepresented by L².

A compound of formula XVI is conveniently used in which L² represents aphenoxy group which may if desired carry up to 5 substituents,preferably up to 2 substituents, selected from halogeno, nitro andcyano. The reaction may be conveniently effected under conditions asdescribed for process (b) hereinbefore.

The compounds of formula XVI and salts thereof as hereinbefore definedmay for example be prepared by deprotecting a compound of the formulaXVII:

(wherein X¹ and L² are as hereinbefore defined and P¹ represents aphenolic hydroxy protecting group). The choice of phenolic hydroxyprotecting group P¹ is within the standard knowledge of an organicchemist, for example those included in standard texts such as“Protective Groups in Organic Synthesis” T. W. Greene and R. G. M. Wuts,2nd Ed. Wiley 1991, including ethers (for example, methyl,methoxymethyl, allyl and benzyl and benzyl substituted with up to twosubstituents selected from C₁₋₄alkoxy and nitro), silyl ethers (forexample, t-butyldiphenylsilyl and t-butyldimethylsilyl), esters (forexample, acetate and benzoate) and carbonates (for example, methyl andbenzyl and benzyl substituted with up to two substituents selected fromC₁₋₄alkoxy and nitro). Deprotection may be effected by techniques wellknown in the literature, for example where P¹ represents a benzyl groupdeprotection may be effected by hydrogenolysis or by treatment withtrifluoroacetic acid.

The removal of such a phenolic hydroxy protecting group may be effectedby any of the procedures known for such a transformation, includingthose reaction conditions indicated in standard texts such as thatindicated hereinbefore, or by a related procedure. The reactionconditions preferably being such that the hydroxy derivative is producedwithout unwanted reactions at other sites within the starting or productcompounds. For example, where the protecting group P′ is acetate, thetransformation may conveniently be effected by treatment of thequinazoline derivative with a base as defined hereinbefore and includingammonia, and its mono and di-alkylated derivatives, preferably in thepresence of a protic solvent or co-solvent such as water or an alcohol,for example methanol or ethanol. Such a reaction can be effected in thepresence of an additional inert solvent or diluent as definedhereinbefore and at a temperature in the range 0 to 50° C., convenientlyat about 20° C.

One compound of formula III may if desired be converted into anothercompound of formula III in which the moiety L¹ is different. Thus forexample a compound of formula III in which L¹ is other than halogeno,for example optionally substituted phenoxy, may be converted to acompound of formula III in which L¹ is halogeno by hydrolysis of acompound of formula III (in which L¹ is other than halogeno) to yield acompound of formula X as hereinbefore defined, followed by introductionof halide to the compound of formula X, thus obtained as hereinbeforedefined, to yield a compound of formula III in which L¹ representshalogeno.

(ii) Compounds of the formula V as hereinbefore defined and saltsthereof may be made by deprotecting the compound of formula XVIII:

(wherein R¹, P¹, X¹ and m are as hereinbefore defined) by a process forexample as described in (i) above.

Compounds of the formula XVIII and salts thereof may be made by reactingcompounds of the formulae XVII and IV as hereinbefore defined, under theconditions described in (a) hereinbefore, to give a compound of theformula XVIII or salt thereof.

(iii) Compounds of the formula VII and salts thereof as hereinbeforedefined may be made by reacting a compound of the formula XIX:

(wherein L¹ is as hereinbefore defined, and L¹ in the 4- and 7-positionsmay be the same or different) with a compound of formual IV ashereinbefore defined, the reaction for example being effected by aprocess as described in (a) above.

(iv) A compound of the formula IX may be prepared by the reaction of acompound of the formula V as defined hereinbefore with a compound of theformula XX:

R⁴-L¹   (XX)

wherein R⁴ and L¹ are as defined hereinbefore under the conditionsdescribed in (b) hereinbefore to give a compound of the formula IX orsalt thereof. The reaction is preferably effected in the presence of abase (as defined hereinbefore in process (a)) and advantageously in thepresence of an inert solvent or diluent (as defined hereinbefore inprocess (a)), advantageously at a temperature in the range, for example10 to 150° C., conveniently in the range 20-50° C.

When a pharmaceutically acceptable salt of a compound of the formula Iis required, it may be obtained, for example, by reaction of saidcompound with, for example, an acid using a conventional procedure, theacid having a pharmaceutically acceptable anion, or it may be obtainedby reaction of said compound with a base by a conventional procedure.

The identification of compounds which potently inhibit the tyrosinekinase activity associated with the VEGF receptors such as Flt and/orKDR and which inhibit angiogenesis and/or increased vascularpermeability is desirable and is the subject of the present invention.These properties may be assessed, for example, using one or more of theprocedures set out below:

(a) In Vitro Receptor Tyrosine Kinase Inhibition Test

This assay determines the ability of a test compound to inhibit tyrosinekinase activity. DNA encoding VEGF or epidermal growth factor (EGF)receptor cytoplasmic domains may be obtained by total gene synthesis(Edwards M, International Biotechnology Lab 5(3), 19-25, 1987) or bycloning. These may then be expressed in a suitable expression system toobtain 30 polypeptide with tyrosine kinase activity. For example VEGFand EGF receptor cytoplasmic domains, which were obtained by expressionof recombinant protein in insect cells, were found to display intrinsictyrosine kinase activity. In the case of the VEGF receptor Flt (Genbankaccession number X51602), a 1.7 kb DNA fragment encoding most of thecytoplasmic domain, commencing with methionine 783 and including thetermination codon, described by Shibuya et al (Oncogene, 1990, 5:519-524), was isolated from cDNA and cloned into a baculovirustransplacement vector (for example pAcYM1 (see The BaculovirusExpression System: A Laboratory Guide, L. A. King and R. D. Possee,Chapman and Hall, 1992) or pAc360 or pBlueBacHis (available fromInvitrogen Corporation)). This recombinant construct was co-transfectedinto insect cells (for example Spodoptera frugiperda 21(Sf21)) withviral DNA (eg Pharmingen BaculoGold) to prepare recombinant baculovirus.(Details of the methods for the assembly of recombinant DNA moleculesand the preparation and use of recombinant baculovirus can be found instandard texts for example Sambrook et al, 1989, Molecular cloning—ALaboratory Manual, 2nd edition, Cold Spring Harbour Laboratory Press andO'Reilly et al, 1992, Baculovirus Expression Vectors - A LaboratoryManual, W. H. Freeman and Co, New York). For other tyrosine kinases foruse in assays, cytoplasmic fragments starting from methionine 806 (KDR,Genbank accession number L04947) and methionine 668 (EGF receptor,Genbank accession number X00588) may be cloned and expressed in asimilar manner.

For expression of cFlt tyrosine kinase activity, Sf21 cells wereinfected with plaque-pure cFlt recombinant virus at a multiplicity ofinfection of 3 and harvested 48 hours later. Harvested cells were washedwith ice cold phosphate buffered saline solution (PBS) (10 mM sodiumphosphate pH7.4, 138 mM sodium chloride, 2.7 mM potassium chloride) thenresuspended in ice cold HNTG/PMSF (20 mM Hepes pH7.5, 150 mM sodiumchloride, 10% v/v glycerol, 1% v/v Triton X100, 1.5 mM magnesiumchloride, 1 mM ethylene glycol-bis(βaminoethyl ether)N,N,N′,N′-tetraacetic acid (EGTA), 1 mM PMSF (phenylmethylsulphonylfluoride); the PMSF is added just before use from a freshly-prepared 100mM solution in methanol) using lml HNTG/PMSF per 10 million cells. Thesuspension was centrifuged for 10 minutes at 13,000 rpm at 4° C., thesupernatant (enzyme stock) was removed and stored in aliquots at −70° C.Each new batch of stock enzyme was titrated in the assay by dilutionwith enzyme diluent (100 mM Hepes pH 7.4, 0.2 mM sodium orthovanadate,0.1% v/v Triton X100, 0.2 mM dithiothreitol). For a typical batch, stockenzyme is diluted 1 in 2000 with enzyme diluent and 50 μl of diluteenzyme is used for each assay well.

A stock of substrate solution was prepared from a random copolymercontaining tyrosine, for example Poly (Glu, Ala, Tyr) 6:3:1 (SigmaP3899), stored as 1 mg/ml stock in PBS at −20° C. and diluted 1 in 500with PBS for plate coating.

On the day before the assay 100 μl of diluted substrate solution wasdispensed into all wells of assay plates (Nunc maxisorp 96-wellimmunoplates) which were sealed and left overnight at 4° C.

On the day of the assay the substrate solution was discarded and theassay plate wells were washed once with PBST (PBS containing 0.05% v/vTween 20) and once with 50 mM Hepes pH7.4.

Test compounds were diluted with 10% dimethylsulphoxide (DMSO) and 25 μlof diluted compound was transferred to wells in the washed assay plates.“Total” control wells contained 10% DMSO instead of compound. Twentyfive microlitres of 40mM manganese(II)chloride containing 8 μMadenosine-5′-triphosphate (ATP) was added to all test wells except“blank” control wells which contained manganese(II)chloride without ATP.To start the reactions 50 μl of freshly diluted enzyme was added to eachwell and the plates were incubated at room temperature for 20 minutes.The liquid was then discarded and the wells were washed twice with PBST.One hundred microlitres of mouse IgG anti-phosphotyrosine antibody(Upstate Biotechnology Inc. product 05-321), diluted 1 in 6000 with PBSTcontaining 0.5% w/v bovine serum albumin (BSA), was added to each welland the plates were incubated for 1 hour at room temperature beforediscarding the liquid and washing the wells twice with PBST. One hundredmicrolitres of horse radish peroxidase (HRP)-linked sheep anti-mouse Igantibody (Amersham product NXA 931), diluted 1 in 500 with PBSTcontaining 0.5% w/v BSA, was added and the plates were incubated for 1hour at room temperature before discarding the liquid and washing thewells twice with PBST. One hundred microlitres of2,2′-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) solution,freshly prepared using one 50 mg ABTS tablet (Boehringer 1204 521) in 50ml freshly prepared 50 mM phosphate-citrate buffer pH5.0 +0.03% sodiumperborate (made with 1 phosphate citrate buffer with sodium perborate(PCSB) capsule (Sigma P4922) per 100 ml distilled water), was added toeach well. Plates were then incubated for 20-60 minutes at roomtemperature until the optical density value of the “total” controlwells, measured at 405 nm using a plate reading spectrophotometer, wasapproximately 1.0. “Blank” (no ATP) and “total” (no compound) controlvalues were used to determine the dilution range of test compound whichgave 50% inhibtion of enzyme activity.

(b) In Vitro HUVEC Proliferation Assay

This assay determines the ability of a test compound to inhibit thegrowth factor-stimulated proliferation of human umbilical veinendothelial cells (HUVEC).

HUVEC cells were isolated in MCDB 131 (Gibco BRL)+7.5% v/v foetal calfserum (FCS) and were plated out (at passage 2 to 8), in MCDB 131+2% v/vFCS+3 μg/ml heparin+1 μg/ml hydrocortisone, at a concentration of 1000cells/well in 96 well plates. After a minimum of 4 hours they were dosedwith the appropriate growth factor (i.e. VEGF 3 ng/ml, EGF 3 ng/ml orb-FGF 0.3 ng/ml) and compound. The cultures were then incubated for 4days at 37° C. with 7.5% carbon dioxide. On day 4 the cultures werepulsed with 1 μCi/well of tritiated-thymidine (Amersham product TRA 61)and incubated for 4 hours. The cells were harvested using a 96-wellplate harvester (Tomtek) and then assayed for incorporation of tritiumwith a Beta plate counter. Incorporation of radioactivity into cells,expressed as cpm, was used to measure inhibition of growthfactor-stimulated cell proliferation by compounds.

(c) In Vivo Solid Tumour Disease Model

This test measures the capacity of compounds to inhibit solid tumourgrowth.

CaLu-6 tumour xenografts were established in the flank of female athymicSwiss nu/nu mice, by subcutaneous injection of 1×10⁶ CaLu-6 cells/mousein 1000 of a 50% (v/v) solution of Matrigel in serum free culturemedium. Ten days after cellular implant, mice were allocated to groupsof 8-10, so as to achieve comparable group mean volumes. Tumours weremeasured using vernier calipers and volumes were calculated as:(l×w)×√(l×w)×(π/6) , where 1 is the longest diameter and w the diameterperpendicular to the longest diameter. Test compounds were administeredorally once daily for a minimum of 21 days, and control animals receivedcompound diluent. Tumours were measured twice weekly. The level ofgrowth inhibition was calculated by comparison of the mean tumour volumeof the control group versus the treatment group, and statisticalsignificance determined using a Students' t-test and/or a Mann-WhitneyRank Sum Test. The inhibitory effect of compound treatment wasconsidered significant when p<0.05.

The toxicological profile of compounds of the present invention may beassessed, for example using a rat 14 day study as described hereinafter.

(d) 14 Day Toxicity Test in Rat

This test measures the activity of compounds in increasing the zone ofhypertrophy in the femoral epiphyseal growth plates of the distal femurand proximal tibia, and allows assessment of histopathological changesin other tissues.

Angiogenesis is an essential event in endochondral ossification duringlong bone elongation, and vascular invasion of the growth plate has beensuggested to depend upon VEGF production by hypertrophic chondrocytes.Expansion of the hypertrophic chondrocyte zone and inhibition ofangiogenesis has been demonstrated following treatment with agents whichspecifically sequester VEGF, such as, for example, (i) a soluble VEGFreceptor chimeric protein (Flt-(1-3)-IgG) in mice (Gerber, H-P., Vu, T.H., Ryan, A. M., Kowalski, J.,

Werb, Z. and Ferrara, N. VEGF couples hypertrophic cartilageremodelling, ossification and angiogenesis during endochondral boneformation, Nature Med., 5: 623-628, 1999) and (ii) a recombinanthumanised anti-VEGF monoclonal IgG1 antibody in cynomologus monkey(Ryan, A. M., Eppler, D. B., Hagler, K E., Bruner, R. H., Thomford, P.J., Hall, R. L., Shopp, G. M. and O'Niell, C. A. Preclinical SafetyEvaluation of rhuMAbVEGF, an antiangiogenic humanised monoclonalantibody, Tox. Path., 27: 78-86, 1999).

An inhibitor of VEGF receptor tyrosine kinase activity should thereforealso inhibit vascular invasion of cartilage, and increase the zone ofhypertrophy in the femoral epiphyseal growth plates of the distal femurand proximal tibia in growing animals.

Compounds were initially formulated by suspension in a 1% (v/v) solutionof polyoxyethylene (20) sorbitan mono-oleate in deionised water, byball-milling at 4° C. overnight (at least 15 hours). Compounds werere-suspended by agitation immediately prior to dosing. Young AlderleyPark rats (Wistar derived, 135-150 g in weight, 4 to 8 weeks of age, 5-6per group) were dosed once-daily by oral gavage for 14 consecutive dayswith compound (at 0.25 ml/100 g body weight) or vehicle. On day 15animals were humanely terminated using a rising concentration of carbondioxide, and a post-mortem performed. A range of tissues, which includedfemoro-tibial joints, were collected and processed by standardhistological techniques to produce paraffin wax sections. Histologicalsections were stained with haematoxylin and eosin and examined by lightmicroscopy for histopathology. The femoral epiphyseal growth plate areasof the distal femur and proximal tibia were measured in sections offemur and tibia using morphometric image analysis. The increase in thezone of hypertrophy was determined by comparison of the mean epiphysealgrowth plate area of the control group versus the treatment group, andstatistical significance determined using a one-tailed Students' t-test.The inhibitory effect of compound treatment was considered significantwhen p<0.05.

Although the pharmacological properties of the compounds of Formula Ivary with structural change, in general, activity possessed by compoundsof the Formula I, may be demonstrated at the following concentrations ordoses in one or more of the above tests (a), (b), (c) and (d):

Test (a):—IC₅₀ in the range, for example, <5 μM;

Test (b):—IC₅₀ in the range, for example, 0.001-5 μM;

Test (c):—activity in the range, for example, 0.1-100 mg/kg;

Test (d):—activity in the range, for example, 0.1-100 mg/kg.

According to one aspect of the present invention compounds of Formula I,assessed in the 14 day toxicity test in rat, have a beneficialtoxicological profile over other compounds within the scope ofInternational Patent Application Publication No. WO 98/13354.

According to another aspect of the present invention compounds ofFormula I, assessed in the 14 day toxicity test in rat, have abeneficial toxicological profile over other compounds within the scopeof International Patent Application Publication No. WO 97/30035.

Although the pharmacological properties of the compounds of Formula Ivary with structural change and between species, at doses in the rat,preferably at doses less than or equal to 150 mg/kg, more preferably atdoses less than or equal to 100 mg/kg, especially at doses less than orequal to 50 mg/kg, compounds of Formula I which produce a statisticallysignificant increase in the femoral epiphyseal growth plate area of thedistal femur and/or proximal tibia, produce no unacceptablehistopathological changes in other tissues in tests (d) that we haveconducted.

Thus by way of example, the compound4-(4-bromo-2-fluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline,(Example 2), tested according to (a), (b), (c) and (d) above gave thefollowing results:

-   (a) Flt—IC₅₀ of 1.6 μM

KDR—IC₅₀ of 0.04 μM

EGFR—IC₅₀ of 0.5 μM

-   (b) VEGF—IC₅₀ of 0.06 μM

EGF—IC₅₀ of 0.17 μM

Basal—IC₅₀ of >3 μM

-   (c) 78% inhibition of tumour growth at 50 mg/kg; p<0.001    (Mann-Whitney Rank Sum Test);-   (d) 75% increase in epiphyseal growth plate hypertrophy at 100    mg/kg/day in female rats; p<0.001 (one-tailed Students' t-test).

According to a further aspect of the invention there is provided apharmaceutical composition which comprises a compound of the formula Ias defined hereinbefore or a pharmaceutically acceptable salt thereof,in association with a pharmaceutically acceptable excipient or carrier.

The composition may be in a form suitable for oral administration, (forexample as tablets, lozenges, hard or soft capsules, aqueous or oilysuspensions, emulsions, dispersible powders or granules, syrups orelixirs), for administration by inhalation (for example as a finelydivided powder or a liquid aerosol), for administration by insufflation(for example as a finely divided powder), for parenteral injection (forexample as a sterile solution, suspension or emulsion for intravenous,subcutaneous, intramuscular, intravascular or infusion dosing), fortopical administration (for example as creams, ointments, gels, oraqueous or oily solutions or suspensions), or for rectal administration(for example as a suppository). In general the above compositions may beprepared in a conventional manner using conventional excipients.

The compositions of the present invention are advantageously presentedin unit dosage form. The compound will normally be administered to awarm-blooded animal at a unit dose within the range 5-5000 mg per squaremetre body area of the animal, i.e. approximately 0.1-100 mg/kg. A unitdose in the range, for example, 1-100 mg/kg, preferably 1-50 mg/kg isenvisaged and this normally provides a therapeutically-effective dose. Aunit dose form such as a tablet or capsule will usually contain, forexample 1-250 mg of active ingredient.

According to a further aspect of the present invention there is provideda compound of the formula I or a pharmaceutically acceptable saltthereof as defined hereinbefore for use in a method of treatment of thehuman or animal body by therapy.

We have found that compounds of the present invention inhibit VEGFreceptor tyrosine kinase activity and are therefore of interest fortheir antiangiogenic effects and/or their ability to cause a reductionin vascular permeability.

A further feature of the present invention is a compound of formula I,or a pharmaceutically acceptable salt thereof, for use as a medicament,conveniently a compound of formula I, or a pharmaceutically acceptablesalt thereof, for use as a medicament for producing an antiangiogenicand/or vascular permeability reducing effect in a warm-blooded animalsuch as a human being.

Thus according to a further aspect of the invention there is providedthe use of a compound of the formula I, or a pharmaceutically acceptablesalt thereof in the manufacture of a medicament for use in theproduction of an antiangiogenic and/or vascular permeability reducingeffect in a warm-blooded animal such as a human being.

According to a further feature of the invention there is provided amethod for producing an antiangiogenic and/or vascular permeabilityreducing effect in a warm-blooded animal, such as a human being, in needof such treatment which comprises administering to said animal aneffective amount of a compound of formula I or a pharmaceuticallyacceptable salt thereof as defined hereinbefore.

As stated above the size of the dose required for the therapeutic orprophylactic treatment of a particular disease state will necessarily bevaried depending on the host treated, the route of administration andthe severity of the illness being treated. Preferably a daily dose inthe range of 1-50 mg/kg is employed. However the daily dose willnecessarily be varied depending upon the host treated, the particularroute of administration, and the severity of the illness being treated.Accordingly the optimum dosage may be determined by the practitioner whois treating any particular patient.

The antiangiogenic and/or vascular permeability reducing treatmentdefined hereinbefore may be applied as a sole therapy or may involve, inaddition to a compound of the invention, one or more other substancesand/or treatments. Such conjoint treatment may be achieved by way of thesimultaneous, sequential or separate administration of the individualcomponents of the treatment. In the field of medical oncology it isnormal practice to use a combination of different forms of treatment totreat each patient with cancer. In medical oncology the othercomponent(s) of such conjoint treatment in addition to theantiangiogenic and/or vascular permeability reducing treatment definedhereinbefore may be: surgery, radiotherapy or chemotherapy. Suchchemotherapy may cover five main categories of therapeutic agent:

(i) other antiangiogenic agents that work by different mechanisms fromthose defined hereinbefore (for example linomide, inhibitors of integrinαvβ3 function, angiostatin, endostatin, razoxin, thalidomide) andincluding vascular targeting agents (for example combretastatinphosphate and the vascular damaging agents described in InternationalPatent Application Publication No. WO 99/02166 the entire disclosure ofwhich document is incorporated herein by reference, (for exampleN-acetylcolchinol-O-phosphate));

(ii) cytostatic agents such as antioestrogens (for exampletamoxifen,toremifene, raloxifene, droloxifene, iodoxyfene), progestogens(for example megestrol acetate), aromatase inhibitors (for exampleanastrozole, letrazole, vorazole, exemestane), antiprogestogens,antiandrogens (for example flutamide, nilutamide, bicalutamide,cyproterone acetate), LHRH agonists and antagonists (for examplegoserelin acetate, luprolide, abarelix), inhibitors of testosterone5α-dihydroreductase (for example finasteride), anti-invasion agents (forexample metalloproteinase inhibitors like marimastat and inhibitors ofurokinase plasminogen activator receptor function) and inhibitors ofgrowth factor function, (such growth factors include for exampleplatelet derived growth factor and hepatocyte growth factor suchinhibitors include growth factor antibodies, growth factor receptorantibodies, tyrosine kinase inhibitors and serine/threonine kinaseinhibitors);

(iii) biological response modifiers (for example interferon);

(iv) antibodies (for example edrecolomab); and

(v) antiproliferative/antineoplastic drugs and combinations thereof, asused in medical oncology, such as antimetabolites (for exampleantifolates like methotrexate, fluoropyrimidines like 5-fluorouracil,purine and adenosine analogues, cytosine arabinoside); antitumourantibiotics (for example anthracyclines like doxorubicin, daunomycin,epirubicin and idarubicin, mitomycin-C, dactinomycin, mithramycin);platinum derivatives (for example cisplatin, carboplatin); alkylatingagents (for example nitrogen mustard, melphalan, chlorambucil,busulphan, cyclophosphamide, ifosfamide, nitrosoureas, thiotepa);antimitotic agents (for example vinca alkaloids like vincristine andtaxoids like taxol, taxotere); enzymes (for example asparaginase);thymidylate synthase inhibitors (for example raltitrexed); topoisomeraseinhibitors (for example epipodophyllotoxins like etoposide andteniposide, amsacrine, topotecan, irinotecan).

For example such conjoint treatment may be achieved by way of thesimultaneous, sequential or separate administration of a compound offormula 1 as defined hereinbefore such as4-(4-bromo-2-fluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazolineor a salt thereof especially a hydrochloride salt thereof, and avascular targeting agent described in WO 99/02166 such asN-acetylcolchinol-O-phosphate (Exampe 1 of WO 99/02166).

As stated above the compounds defined in the present invention are ofinterest for their antiangiogenic and/or vascular permeability reducingeffects. Such compounds of the invention are expected to be useful in awide range of disease states including cancer, diabetes, psoriasis,rheumatoid arthritis, Kaposi's sarcoma, haemangioma, acute and chronicnephropathies, atheroma, arterial restenosis, autoimmune diseases, acuteinflammation, excessive scar formation and adhesions, endometriosis,dysfunctional uterine bleeding and ocular diseases with retinal vesselproliferation. In particular such compounds of the invention areexpected to slow advantageously the growth of primary and recurrentsolid tumours of, for example, the colon, breast, prostate, lungs andskin. More particularly such compounds of the invention are expected toinhibit the growth of those primary and recurrent solid tumours whichare associated with VEGF especially those tumours which aresignificantly dependent on VEGF for their growth and spread, includingfor example, certain tumours of the colon, breast, prostate, lung, vulvaand skin.

In another aspect of the present invention compounds of Formula I areexpected to inhibit the growth of those primary and recurrent solidtumours which are associated with EGF especially those tumours which aresignificantly dependent on EGF for their growth and spread.

In another aspect of the present invention compounds of Formula I areexpected to inhibit the growth of those primary and recurrent solidtumours which are associated with both VEGF and EGF especially thosetumours which are significantly dependent on VEGF and EGF for theirgrowth and spread.

In addition to their use in therapeutic medicine, the compounds offormula I and their pharmaceutically acceptable salts are also useful aspharmacological tools in the development and standardisation of in vitroand in vivo test systems for the evaluation of the effects of inhibitorsof VEGF receptor tyrosine kinase activity in laboratory animals such ascats, dogs, rabbits, monkeys, rats and mice, as part of the search fornew therapeutic agents.

It is to be understood that where the term “ether” is used anywhere inthis specification it refers to diethyl ether.

The invention will now be illustrated, but not limited, by the followingExamples in which, unless otherwise stated:

(i) evaporations were carried out by rotary evaporation in vacuo andwork-up procedures were carried out after removal of residual solidssuch as drying agents by filtration;

(ii) operations were carried out at ambient temperature, that is in therange 18-25° C. and under an atmosphere of an inert gas such as argon;

(iii) column chromatography (by the flash procedure) and medium pressureliquid chromatography (MPLC) were performed on Merck Kieselgel silica(Art. 9385) or Merck Lichroprep RP-18 (Art. 9303) reversed-phase silicaobtained from E. Merck, Darmstadt, Germany;

(iv) yields are given for illustration only and are not necessarily themaximum attainable;

(v) melting points are uncorrected and were determined using a MettlerSP62 automatic melting point apparatus, an oil-bath apparatus or aKoffler hot plate apparatus.

(vi) the structures of the end-products of the formula I were confirmedby nuclear (generally proton) magnetic resonance (NMR) and mass spectraltechniques; proton magnetic resonance chemical shift values weremeasured on the delta scale and peak multiplicities are shown asfollows: s, singlet; d, doublet; t, triplet; m, multiplet; br, broad; q,quartet; NMR spectra were run on a 400 MHz machine at 24° C.

(vii) intermediates were not generally fully characterised and puritywas assessed by thin layer chromatography (TLC), high-performance liquidchromatography (HPLC), infra-red (IR) or NMR analysis;

(viii) the following abbreviations have been used:

-   -   DMF N,N-dimethylformamide

DMSO dimethylsulphoxide

-   -   THF tetrahydrofuran    -   TFA trifluoroacetic acid    -   NMP 1-methyl-2-pyrrolidinone.]

EXAMPLE 1

TFA (3 ml) was added to a suspension of4-(4-bromo-2-fluoroanilino)-7-(1-(tert-butoxycarbonyl)piperidin-4-ylmethoxy)-6-methoxyquinazoline(673 mg, 1.2 mmol) in methylene chloride (10 ml). After stirring for 1hour at ambient temperature, the volatiles were removed under vacuum.The residue was triturated with a mixture of water/ether. The organiclayer was separated. The aqueous layer was washed again with ether. Theaqueous layer was adjusted to pH10 with 2N aqueous sodium hydroxide. Theaqueous layer was extracted with methylene chloride. The organic layerwas dried (MgSO₄) and the solvent was removed under vacuum. The solidwas triturated with a mixture ether/petroleum ether (1/1), filtered,washed with ether and dried under vacuum to give4-(4-bromo-2-fluoroanilino)-6-methoxy-7-(piperidin-4-ylmethoxy)quinazoline(390 mg, 70.5%).

MS—ESI: 461-463 [MH]⁺

¹H NMR Spectrum: (DMSOd₆) 1.13-1.3 (m, 2H), 1.75 (d, 2H), 1.87-2.0 (m,1H), 2.5 (d, 2H), 3.0 (d, 2H), 3.96 (s, 3H), 3.98 (d, 2H), 7.2 (s, 1H),7.5 (dd, 1H), 7.55 (t, 1H), 7.68 (dd, 1H), 7.80 (s, 1H), 8.36 (s, 1H),9.55 (br s, 1H)

Elemental analysis: Found C 54.5 H 4.9 N 12.1 C₂₁H₂₂N₄O₂BrF Requires C54.7 H 4.8 N 12.1%

The starting material was prepared as follows:

A solution of 7-benzyloxy-4-chloro-6-methoxyquinazoline hydrochloride(8.35 g, 27.8 mmol), (prepared, for example, as described in WO97/22596, Example 1), and 4-bromo-2-fluoroaniline (5.65 g, 29.7 mmol) in2-propanol (200 ml) was heated at reflux for 4 hours. The resultingprecipitate was collected by filtration, washed with 2-propanol and thenether and dried under vacuum to give7-benzyloxy-4-(4-bromo-2-fluoroanilino)-6-methoxyquinazolinehydrochloride (9.46 g, 78%).

¹H NMR Spectrum: (DMSO d₆; CD₃COOD) 4.0(s, 3H); 5.37(s, 2H); 7.35-7.5(m,4H); 7.52-7.62(m, 4H); 7.8(d, 1H); 8.14(9s, 1H); 8.79(s, 1H)

MS—ESI: 456 [MH]⁺

Elemental analysis: Found C 54.0 H 3.7 N 8.7 C₂₂H₁₇N₃O₂BrF 0.9HClRequires C 54.2 H 3.7 N 8.6%

A solution of7-benzyloxy-4-(4-bromo-2-fluoroanilino)-6-methoxyquinazolinehydrochloride (9.4 g, 19.1 mmol) in TFA (90 ml) was heated at reflux for50 minutes. The mixture was allowed to cool and was poured on to ice.The resulting precipitate was collected by filtration and dissolved inmethanol (70 ml). The solution was adjusted to pH9-10 with concentratedaqueous ammonia solution. The mixture was concentrated to half initialvolume by evaporation. The resulting precipitate was collected byfiltration, washed with water and then ether, and dried under vacuum togive 4-(4-bromo-2-fluoroanilino)-7-hydroxy-6-methoxyquinazoline (5.66 g,82%).

¹H NMR Spectrum: (DMSO d₆; CD₃COOD) 3.95(s, 3H); 7.09(s, 1H); 7.48(s,1H); 7.54(t, 1H); 7.64(d, 1H); 7.79(s, 1H); 8.31(s, 1H)

MS—ESI: 366 [MH]⁺

Elemental analysis: Found C 49.5 H 3.1 N 11.3 C₁₅H₁₁N₃O₂BrF Requires C49.5 H 3.0 N 11.5%

While maintaining the temperature in the range 0-5° C., a solution ofdi-cert-butyl dicarbonate (41.7 g, 0.19 mol) in ethyl acetate (75 ml)was added in portions to a solution of ethyl 4-piperidinecarboxylate (30g, 0.19 mol) in ethyl acetate (150 ml) cooled at 5° C. After stirringfor 48 hours at ambient temperature, the mixture was poured onto water(300 ml). The organic layer was separated, washed successively withwater (200 ml), 0.1 N aqueous hydrochloric acid (200m1), saturatedsodium hydrogen carbonate (200 ml) and brine (200 ml), dried (MgSO₄) andevaporated to give ethyl4-(1-(tert-butoxycarbonyl)piperidine)carboxylate (48 g, 98%).

¹H NMR Spectrum: (CDCl₃) 1.25(t, 3H); 1.45(s, 911); 1.55-1.70(m, 2H);1.8-2.0(d, 2H); 2.35-2.5(m, 1H); 2.7-2.95(t, 2H); 3.9-4.1(br s, 2H);4.15 (q, 2H)

A solution of 1M lithium aluminium hydride in THF (133 ml, 0.133 mol)was added in portions to a solution of ethyl4-(1-(tert-butoxycarbonyl)piperidine)carboxylate (48 g, 0.19 mol) in dryTHF (180 ml) cooled at 0° C. After stirring at 0° C. for 2 hours, water(30 ml) was added followed by 2N sodium hydroxide (10 ml). Theprecipitate was removed by filtration through diatomaceous earth andwashed with ethyl acetate. The filtrate was washed with water, brine,dried (MgSO₄) and evaporated to give1-(tert-butoxycarbonyl)-4-hydroxymethylpiperidine (36.3 g, 89%).

MS (EI): 215 [M.]+

¹H NMR Spectrum: (CDCl₃) 1.05-1.2(m, 2H); 1.35-1.55(m, 10H); 1.6-1.8(m,2H); 2.6-2.8(t, 2H); 3.4-3.6(t, 2H); 4.0-4.2(br s, 2H)

1,4-Diazabicyclo[2.2.2]octane (42.4 g, 0.378 mol) was added to asolution of 1-(tert-butoxycarbonyl)-4-hydroxymethylpiperidine (52.5 g,0.244 mol) in tert-butyl methyl ether (525 ml). After stirring for 15minutes at ambient temperature, the mixture was cooled to 5° C. and asolution of toluene sulphonyl chloride (62.8 g, 0.33 mmol) in tert-butylmethyl ether (525 ml) was added in portions over 2 hours whilemaintaining the temperature at 0° C. After stirring for 1 hour atambient temperature, petroleum ether (1 l) was added. The precipitatewas removed by filtration. The filtrate was evaporated to give a solid.The solid was dissolved in ether and washed successively with 0.5Naqueous hydrochloric acid (2×500 ml), water, saturated sodium hydrogencarbonate and brine, dried (MgSO₄) and evaporated to give1-(tert-butoxycarbonyl)-4-(4-methylphenylsulphonyloxymethyl)piperidine(76.7 g, 85%). MS (ESI): 392 [MNa]⁺

¹H NMR Spectrum: (CDCl₃) 1.0-1.2(m, 2H); 1.45(s, 9H); 1.65(d, 2H);1.75-1.9(m, 2H); 2.45(s, 3H); 2.55-2.75(m, 2H); 3.85(d, 1H); 4.0-4.2(brs, 2H); 7.35(d, 2H); 7.8(d, 2H)

Potassium carbonate (414 mg, 3 mmol) was added to a suspension of4-(4-bromo-2-fluoroanilino)-7-hydroxy-6-methoxyquinazoline (546 mg, 1.5mmol) in DMF (5 ml). After stirring for 10 minutes at ambienttemperature,1-(tert-butoxycarbonyl)-4-(4-methylphenylsulphonyloxymethyl)piperidine(636 mg, 1.72 mmol) was added and the mixture was heated at 95° C. for 2hours. After cooling, the mixture was poured onto cooled water (20 ml).The precipitate was collected by filtration, washed with water, anddried under vacuum to give4-(4-bromo-2-fluoroanilino)-7-(1-(tert-butoxycarbonyl)piperidin-4-ylmethoxy)-6-methoxyquinazoline(665 mg, 79%).

MS—ESI: 561-563 [MH]⁺

¹H NMR Spectrum: (DMSO d₆) 1.15-1.3 (m, 2H), 1.46 (s, 9H), 1.8 (d, 2H),2.0-2.1 (m, 1H), 2.65-2.9 (m, 2H), 3.95 (s, 3H), 4.02 (br s, 2H), 4.05(d, 2H), 7.2 (s, 1H), 7.48 (d, 1H), 7.55 (t, H), 7.65 (d, 1H), 7.8 (s,1H), 8.35 (s, 1H), 9.55 (br s, 1H)

EXAMPLE 2a

A solution of 37% aqueous formaldehyde (50 μl, 0.6 mmol) followed bysodium cyanoborohydride (23 mg, 0.36 mmol) were added to a solution of4-(4-bromo-2-fluoroanilino)-6-methoxy-7-(piperidin-4-ylmethoxy)quinazoline(139 mg, 0.3 mmol), (prepared as described in Example 1), in a mixtureof THF/methanol (1.4 ml/1.4 ml). After stirring for 1 hour at ambienttemperature, water was added and the volatiles were removed undervacuum. The residue was triturated with water, filtered, washed withwater, and dried under vacuum. The solid was purified by chromatographyon neutral alumina eluting with methylene chloride followed by methylenechloride/ethyl acetate (1/1) followed by methylene chloride/ethylacetate/methanol (50/45/5). The fractions containing the expectedproduct were evaporated under vacuum. The resulting white solid wasdissolved in methylene chloride/methanol (3 ml/3 ml) and 3N hydrogenchloride in ether (0.5 ml) was added. The volatiles were removed undervacuum. The solid was triturated with ether, filtered, washed with etherand dried under vacuum to give4-(4-bromo-2-fluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazolinehydrochloride (120 mg, 69%).

MS—ESI: 475-477 [MH]⁺

The NMR spectrum of the protonated form of4-(4-bromo-2-fluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazolinehydrochloride shows the presence of 2 forms A and B in a ratio A:B ofapproximately 9:1.

¹H NMR Spectrum: (DMSOd₆; CF₃COOD) 1.55-1.7 (m, form A 2H); 1.85-2.0 (m,form B 4H); 2.03 (d, form A 2H); 2.08-2.14 (br s, form A 1H); 2.31-2.38(br s, form B 1H); 2.79 (s, form A 3H); 2.82 (s, form B 3H); 3.03 (t,form A 2H); 3.21 (br s, form B 2H); 3.30 (br s, form B 2H); 3.52 (d,form A 2H); 4.02 (s, 3H); 4.12 (d, form A 2H); 4.30 (d, form B 2H); 7.41(s, 1H); 7.5-7.65 (m, 2H); 7.81 (d, 1H); 8.20 (s, 1H); 8.88 (s, 1H)

Elemental analysis: Found C 46.0 H 5.2 N 9.6 C₂₂H₂₄N₄O₂BrF 0.3H₂ORequires C 45.8 H 4.8 N 9.7% 2.65HCl

Example 2b

37% Aqueous formaldehyde (3.5 ml, 42 mmol) was added to a solution of4-(4-bromo-2-fluoroanilino)-7-(1-(tert-butoxycarbonyl)piperidin-4-ylmethoxy)-6-methoxyquinazoline(3.49 g, 6.22 mmol), (prepared as described for the starting material inExample 1), in formic acid (35 ml). After heating at 95° C. for 4 hoursthe volatiles were removed under vacuum. The residue was suspended inwater and the mixture was adjusted to pH10.5 by slow addition of asolution of 2N sodium hydroxide. The suspension was extracted with ethylacetate. The organic layer was washed with brine, dried MgSO₄ andevaporated to give4-(4-bromo-2-fluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline(2.61 g, 88%).

MS—ESI: 475-477 [MH]⁺

¹H NMR Spectrum: (DMSOd₆) 1.3-1.45 (m, 2H), 1.8 (d, 2H), 1.7-1.9 (m,1H), 1.95 (t, 2H), 2.2 (s, 3H), 2.85 (d, 2H), 3.96 (s, 3H), 4.05 (d,2H), 7.19 (s, 1H), 7.5 (d, 1H), 7.55 (t, 1H), 7.67 (d, 1H), 7.81 (s,1H), 8.37 (s, 1H), 9.54 (s, 1H)

Elemental analysis: Found C 55.4 H 5.1 N 11.6 C₂₂H₂₄N₄O₂BrF Requires C55.6 H 5.1 N 11.8%

EXAM PLE 2c

A suspension of4-chloro-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline (200 mg,0.62 mmol) and 4-bromo-2-fluoroaniline (142 mg, 0.74 mmol) inisopropanol (3 ml) containing 6N hydrogen chloride in isopropanol (110μl, 0.68 ml) was heated at reflux for 1.5 hours. After cooling, theprecipitate was collected by filtration, washed with isopropanolfollowed by ether and dried under vacuum to give4-(4-bromo-2-fluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazolinehydrochloride (304 mg, 90%).

Elemental analysis: Found C 47.9 H 4.9 N 10.0 C₂₂H₂₄N₄O₂BrF 0.5H₂ORequires C 48.2 H 5.0 N 10.1% 1.8HCl0.08 isopropanol

The NMR spectrum of the protonated form of4-(4-bromo-2-fluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazolinehydrochloride shows the presence of two forms A and B in a ratio A:B ofapproximately 9:1.

¹H NMR Spectrum: (DMSOd₆) 1.6-1.78 (m, form A 2H); 1.81-1.93 (br s, formB 4H); 1.94-2.07 (d, form A 2H); 2.08-2.23 (br s, form A 1H); 2.29-2.37(br s, form B 1H); 2.73 (d, form A 3H); 2.77 (d, form B 3H); 2.93-3.10(q, form A 2H); 3.21 (br s, form B 2H); 3.27 (br s, form B 2H);3.42-3.48 (d, form A 2H); 4.04 (s, 3H); 4.10 (d, form A 2H); 4.29 (d,form B 2H); 7.49 (s, 1H); 7.53-7.61 (m, 2H); 7.78 (d, 1H); 8.47 (s, 1H);8.81 (s, 1H); 10.48 (br s, form A 1H); 10.79 (br s, form B 1H); 11.90(br s, 1H)

For another NMR reading, some solid potassium carbonate was added intothe DMSO solution of the4-(4-bromo-2-fluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazolinehydrochloride described above, in order to release the free base in theNMR tube. The NMR spectrum was then recorded again and showed only oneform as described below:

¹H-NMR Spectrum: (DMSO d₆; solid potassium carbonate) 1.3-1.45 (m, 2H);1.75 (d, 2H); 1.7-1.9(m, 1H); 1.89 (t, 2H); 2.18 (s, 3H); 2.8 (d, 2H);3.98 (s, 3H); 4.0 (d, 2H); 7.2 (s, 1H); 7.48 (d, 1H); 7.55 (t, 1H); 7.68(d, 1H); 7.8 (s, 1H); 8.35 (s, 1H); 9.75 (s, 1H)

A sample of4-(4-bromo-2-fluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline(free base) was generated from the4-(4-bromo-2-fluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazolinehydrochloride, (prepared as described above), as follows:

4-(4-Bromo-2-fluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxyquinazolinehydrochloride (50 mg) was suspended in methylene chloride (2 ml) and waswashed with saturated sodium hydrogen carbonate. The methylene chloridesolution was dried (MgSO₄) and the volatiles were removed by evaporationto give4-(4-bromo-2-fluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline(free base). The NMR of the free base so generated shows only one formas described below:

¹H NMR Spectrum: (DMSOd₆) 1.3-1.45 (m, 2H); 1.76 (d, 2H); 1.7-1.9(m,1H); 1.9 (t, 2H); 2.19 (s, 3H); 2.8 (d, 2H); 3.95 (s, 3H); 4.02 (d, 2H);7.2 (s, 1H); 7.48 (d, 1H); 7.55 (t, 1H); 7.68 (dd, 1H); 7.8 (s, 1H);8.38 (s, 1H); 9.55(br s, 1H)

For another NMR reading, some CF₃COOD was added into the NMR DMSOsolution of the4-(4-bromo-2-fluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline(free base) described above and the NMR spectrum was recorded again. Thespectrum of the protonated form of the4-(4-bromo-2-fluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazolinetrifluoroacetate salt so obtained shows the presence of two forms A andB in a ratio A:B of approximately 9:1.

¹H NMR Spectrum: (DMSOd₆; CF₃COOD) 1.5-1.7 (m, form A 2H); 1.93 (br s,form B 4H); 2.0-2.1 (d, form A 2H); 2.17 (br s, form A 1H); 2.35 (br s,form B1H); 2.71 (s, form A 3H); 2.73 (s, form B 3H); 2.97-3.09 (t, formA 2H); 3.23 (br s, form B 2H); 3.34 (br s, form B 2H); 3.47-3.57 (d,form A 2H); 4.02 (s, 3H); 4.15 (d, form A 2H); 4.30 (d, form B 2H); 7.2(s, 1H); 7.3-7.5 (m, 2H); 7.6 (d, 1H); 7.9 (s, 1H); 8.7 (s, 1H)

The starting material was prepared as follows:

1-(tert-Butoxycarbonyl)-4-(4-methylphenylsulphonyloxymethyl)piperidine(40 g, 0.11 mol), (prepared as described for the starting material inExample 1), was added to a suspension of ethyl4-hydroxy-3-methoxybenzoate (19.6 g, 0.1 mol) and potassium carbonate(28 g, 0.2 mol) in dry DMF (200 ml). After stirring at 95° C. for 2.5hours, the mixture was cooled to ambient temperature and partitionedbetween water and ethyl acetate/ether. The organic layer was washed withwater, brine, dried (MgSO₄) and evaporated. The resulting oil wascrystallised from petroleum ether and the suspension was storedovernight at 5° C. The solid was collected by filtration, washed withpetroleum ether and dried under vacuum to give ethyl4-(1-(tert-butoxycarbonyl)piperidin-4-ylmethoxy)-3-methoxybenzoate (35g, 89%). m.p. 81-83° C.

MS (ESI): 416 [MNa]⁺

¹H NMR Spectrum: (CDCl₃) 1.2-1.35(m, 2H); 1.4(t, 3H); 1.48(s, 9H);1.8-1.9(d, 2H); 2.0-2.15(m, 2H); 2.75(t, 2H); 3.9(d, 2H); 3.95(s, 3H);4.05-4.25(br s, 211); 4.35(q, 2H); 6.85(d, 1H); 7.55(s, 1H); 7.65(d, 1H)

Elemental analysis: Found C 63.4 H 8.0 N 3.5 C₂₁H₃₁NO₆ 0.3H₂O Requires C63.2 H 8.0 N 3.5%

Formaldehyde (12M, 37% in water, 35 ml, 420 mmol) was added to asolution of ethyl4-(1-(tert-butoxycarbonyl)piperidin-4-ylmethoxy)-3-methoxybenzoate (35g, 89 mmol) in formic acid (35 ml). After stirring at 95° C. for 3hours, the volatiles were removed by evaporation. The residue wasdissolved in methylene chloride and 3M hydrogen chloride in ether (40ml, 120 mmol) was added. After dilution with ether, the mixture wastriturated until a solid was formed. The solid was collected byfiltration, washed with ether and dried under vacuum overnight at 50° C.to give ethyl 3-methoxy-4-(1-methylpiperidin-4-ylmethoxy)benzoate (30.6g, quant.).

MS (ESI): 308 [MH]⁺

¹H NMR Spectrum: (DMSOd₆) 1.29(t, 3H); 1.5-1.7(m, 2H); 1.95(d, 2H);2.0-2.15(br s, 1H); 2.72(s, 3H); 2.9-3.1(m, 2H); 3.35-3.5(br s, 2H);3.85(s, 3H); 3.9-4.05(br s, 2H); 4.3(q, 2H); 7.1(d, 1H); 7.48(s, 1H);7.6(d, 1H)

A solution of ethyl 3-methoxy-4-(1-methylpiperidin-4-ylmethoxy)benzoate(30.6 g, 89 mmol) in methylene chloride (75m1) was cooled to 0-5° C. TFA(37.5m1) was added followed by the dropwise addition over 15 minutes ofa solution of fuming 24N nitric acid (7.42 ml, 178 mmol) in methylenechloride (15 ml). After completion of the addition, the solution wasallowed to warm up and stirred at ambient temperature for 2 hours. Thevolatiles were removed under vacuum and the residue was dissolved inmethylene chloride (50 ml). The solution was cooled to 0-5° C. and etherwas added. The precipitate was collected by filtration, and dried undervacuum at 50° C. The solid was dissolved in methylene chloride (500 ml)and 3M hydrogen chloride in ether (30 ml) was added followed by ether(500 ml). The solid was collected by filtration and dried under vacuumat 50° C. to give ethyl3-methoxy-4-(1-methylpiperidin-4-ylmethoxy)-6-nitrobenzoate (28.4 g,82%).

MS (ESI): 353 [MH]⁺

¹HNMR Spectrum: (DMSOd₆) 1.3(t, 3H); 1.45-1.65(m, 2H); 1.75-2.1(m, 3H);2.75(s, 3H); 2.9-3.05(m, 2H); 3.4-3.5(d, 2H); 3.95(s, 3H); 4.05(d, 2H);4.3(q, 2H); 7.32(s, 1H); 7.66(s, 1H)

A suspension of ethyl3-methoxy-4-(1-methylpiperidin-4-ylmethoxy)-6-nitrobenzoate (3.89 g, 10mmol) in methanol (80 ml) containing 10% platinum on activated carbon(50% wet) (389mg) was hydrogenated at 1.8 atmospheres pressure untiluptake of hydrogen ceased. The mixture was filtered and the filtrate wasevaporated. The residue was dissolved in water (30 ml) and adjusted topH10 with a saturated solution of sodium hydrogen carbonate. The mixturewas diluted with ethyl acetate/ether (1/1) and the organic layer wasseparated. The aqueous layer was further extracted with ethylacetate/ether and the organic layers were combined. The organic layerswere washed with water, brine, dried (MgSO₄), filtered and evaporated.The resulting solid was triturated in a mixture of ether/petroleumether, filtered, washed with petroleum ether and dried under vacuum at60° C. to give ethyl6-amino-3-methoxy-4-(1-methylpiperidin-4-ylmethoxy)benzoate (2.58 g,80%).

m.p. 111-112° C.

MS (ESI): 323 [MH]⁺

¹H NMR Spectrum: (CDCl₃) 1.35(t, 3H); 1.4-1.5(m, 2H); 1.85(m, 3H);1.95(t, 211); 2.29(s, 3H); 2.9(d, 2H); 3.8(s, 3H); 3.85(d, 2H); 4.3(q,2H); 5.55(br s, 2H); 6.13(s, 1H); 7.33(s, 1H)

Elemental analysis: Found C 62.8 H 8.5 N 8.3 C₁₇H₂₆N₂O₄ 0.2H₂O RequiresC 62.6 H 8.2 N 8.6%

A solution of ethyl6-amino-3-methoxy-4-(1-methylpiperidin-4-ylmethoxy)benzoate (16.1 g, 50mmol) in 2-methoxyethanol (160 ml) containing formamidine acetate (5.2g, 50 mmol) was heated at 115° C. for 2 hours. Formamidine acetate (10.4g, 100 mmol) was added in portions every 30 minutes over 4 hours.Heating was prolonged for 30 minutes after the last addition. Aftercooling, the volatiles were removed under vacuum. The solid wasdissolved in ethanol (100 ml) and methylene chloride (50 ml). Theprecipitate was removed by filtration and the filtrate was concentratedto a final volume of 100 ml. The suspension was cooled to 5° C. and thesolid was collected by filtration, washed with cold ethanol followed byether and dried under vacuum overnight at 60° C. to give6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)-3,4-dihydroquinazolin-4-one(12.7g, 70%).

MS (ESI): 304 [MH]⁺

¹H NMR Spectrum: (DMSOd₆) 1.25-1.4(m, 2H); 1.75(d, 2H); 1.9(t, 1H);1.9(s, 3H); 2.16(s, 2H); 2.8(d, 2H); 3.9(s, 3H); 4.0(d, 2H); 7.11(s,1H); 7.44(s, 1H); 7.97(s, 1H)

A solution of6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)-3,4-dihydroquinazolin-4-one(2.8 g, 9.24 mmol) in thionyl chloride (28 ml) containing DMF (280 μl)was heated at reflux at 85° C. for 1 hour. After cooling, the volatileswere removed by evaporation. The precipitate was triturated with ether,filtered, washed with ether and dried under vacuum. The solid wasdissolved in methylene chloride and saturated aqueous sodium hydrogencarbonate was added. The organic layer was separated, washed with water,brine, dried (MgSO₄) and evaporated to give4-chloro-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline (2.9 g,98%).

MS (ESI): 322 [MH]⁺

¹H NMR Spectrum: (DMSOd₆) 1.3-1.5(m, 2H); 1.75-1.9(m, 3H); 2.0(t, 1H);2.25(s, 3H); 2.85(d, 2H); 4.02(s, 3H); 4.12(d, 2H); 7.41(s, 1H); 7.46(s,1H); 8.9(s, 1H)

Alternatively, the6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)-3,4-dihydroquinazolin-4-onecan be prepared as follows:

Sodium hydride (1.44 g of a 60% suspension in mineral oil, 36 mmol) wasadded in portions over 20 minutes to a solution of7-benzyloxy-6-methoxy-3,4-dihydroquinazolin-4-one (8.46 g, 30 mmol),(prepared, for example, as described in WO 97/22596, Example 1), in DMF(70 ml) and the mixture was stirred for 1.5 hours. Chloromethyl pivalate(5.65 g, 37.5 mmol) was added in portions and the mixture stirred for 2hours at ambient temperature.

The mixture was diluted with ethyl acetate (100 ml) and poured ontoice/water (400 ml) and 2N hydrochloric acid (4 ml). The organic layerwas separated and the aqueous layer extracted with ethyl acetate, thecombined extracts were washed with brine, dried (MgSO₄) and the solventremoved by evaporation. The residue was triturated with a mixture ofether and petroleum ether, the solid was collected by filtration anddried under vacuum to give7-benzyloxy-6-methoxy-3-((pivaloyloxy)methyl)-3,4-dihydroquinazolin-4-one(10 g, 84%).

¹H NMR Spectrum: (DMSOd₆) 1.11(s, 9H); 3.89(s, 3H); 5.3(s, 2H); 5.9(s,2H); 7.27(s, 1H); 7.35(m, 1H); 7.47(t, 2H); 7.49(d, 2H); 7.51(s, 1H);8.34(s, 1H)

A mixture of7-benzyloxy-6-methoxy-3-((pivaloyloxy)methyl)-3,4-dihydroquinazolin-4-one(7 g, 17.7 mmol) and 10% palladium-on-charcoal catalyst (700 mg) inethyl acetate (250 ml), DMF (50 ml), methanol (50 ml) and acetic acid(0.7 ml) was stirred under hydrogen at atmospheric pressure for 40minutes. The catalyst was removed by filtration and the solvent removedfrom the filtrate by evaporation. The residue was triturated with ether,collected by filtration and dried under vacuum to give7-hydroxy-6-methoxy-3-((pivaloyloxy)methyl)-3,4-dihydroquinazolin-4-one(4.36 g, 80%).

¹H NMR Spectrum: (DMSOd₆) 1.1(s, 9H); 3.89(s, 3H); 5.89(s, 2H); 7.0(s,1H); 7.48(s, 1H); 8.5(s, 1H)

Triphenyiphosphine (1.7 g, 6.5 mmol) was added under nitrogen to asuspension of7-hydroxy-6-methoxy-3-((pivaloyloxy)methyl)-3,4-dihydroquinazolin-4-one(1.53 g, 5 mmol) in methylene chloride (20 ml), followed by the additionof 1-(tert-butoxycarbonyl)-4-(hydroxymethyl)piperidine (1.29 g, 6 mmol),(prepared as described for the starting material in Example 1), and by asolution of diethyl azodicarboxylate (1.13 g, 6.5 mmol) in methylenechloride (5 ml). After stirring for 30 minutes at ambient temperature,the reaction mixture was poured onto a column of silica and was elutedwith ethyl acetate/petroleum ether (1/1 followed by 6/5, 6/4 and 7/3).Evaporation of the fractions containing the expected product led to anoil that crystallised following trituration with pentane. The solid wascollected by filtration and dried under vacuum to give7-(1-(tert-butoxycarbonyl)piperidin-4-ylmethoxy)-6-methoxy-3-((pivaloyloxy)methyl)-3,4-dihydroquinazolin-4-one(232 g, 92%).

MS—ESI: 526 [MNa]⁺

¹ H NMR Spectrum: (CDCl₃) 1.20 (s, 9H), 1.2-1.35 (m, 2H), 1.43 (s, 9H),1.87 (d, 2H), 2.05-2.2 (m, 1H), 2.75 (t, 2H), 3.96 (d, 2H), 3.97 (s,3H), 4.1-4.25 (br s, 2H), 5.95 (s, 2H), 7.07 (s, 1H), 7.63 (s, 1H), 8.17(s, 1H)

Elemental analysis: Found C 61.8 H 7.5 N 8.3 C₂₆H₃₇N₃O₇ Requires C 62.0H 7.4 N 8.3%

A solution of7-(1-(tert-butoxycarbonyl)piperidin-4-ylmethoxy)-6-methoxy-3-((pivaloyloxy)methyl)-3,4-dihydroquinazolin-4-one(2.32 g, 4.6 mmol) in methylene chloride (23 ml) containing TFA (5 ml)was stirred at ambient temperature for 1 hour. The volatiles wereremoved under vacuum. The residue was partitioned between ethyl acetateand sodium hydrogen carbonate. The organic solvent was removed undervacuum and the residue was filtered. The precipitate was washed withwater, and dried under vacuum. The solid was azeotroped with toluene anddried under vacuum to give6-methoxy-7-(piperidin-4-ylmethoxy)-3-((pivaloyloxy)methyl)-3,4-dihydroquinazolin-4-one(1.7 g, 92%).

MS—ESI: 404 [MH]⁺

¹H NMR Spectrum: (DMSOd₆; CF₃COOD) 1.15 (s, 9H), 1.45-1.6 (m, 2H), 1.95(d, 2H), 2.1-2.25 (m, 1H), 2.95 (t, 2H), 3.35 (d, 2H), 3.95 (s, 3H), 4.1(d, 2H), 5.95 (s, 2H), 7.23 (s, 1H), 7.54 (s, 1H), 8.45 (s, 1H)

A 37% aqueous solution of formaldehyde (501 μl, 6 mmol) followed bysodium cyanoborohydride (228 mg, 3.6 mmol) were added in portions to asolution of6-methoxy-7-(piperidin-4-ylmethoxy)-3-((pivaloyloxy)methyl)-3,4-dihydroquinazolin-4-one(1.21 g, 3 mmol) in a mixture of THF/methanol (10 ml/10 ml). Afterstirring for 30 minutes at ambient temperature, the organic solventswere removed under vacuum and the residue was partitioned betweenmethylene chloride and water. The organic layer was separated, washedwith water and brine, dried (MgSO₄) and the volatiles were removed byevaporation. The residue was triturated with ether and the resultingsolid was collected by filtration, washed with ether and dried undervacuum to give6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)-3-((pivaloyloxy)methyl)-3,4-dihydroquinazolin-4-one(1.02 g, 82%).

MS—ESI: 418 [MH]⁺

¹H NMR Spectrum: (CDCl₃) 1.19 (s, 9H), 1.4-1.55 (m, 2H), 1.9 (d, 2H),2.0 (t, 2H), 1.85-2.1 (m, 1H), 2.3 (s, 3H), 2.92 (d, 2H), 3.96 (s, 3H),3.99 (d, 2H), 5.94 (s, 2H), 7.08 (s, 1H), 7.63 (s, 1H), 8.17 (s, 1H)

A saturated solution of ammonia in methanol (14 ml) was added to asolution of6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)-3-((pivaloyloxy)methyl)-3,4-dihydroquinazolin-4-one(1.38 g, 3.3 mmol) in methanol (5 ml). After stirring for 20 hours atambient temperature, the suspension was diluted with methylene chloride(10 ml). The solution was filtered. The filtrate was evaporated undervacuum and the residue was triturated with ether, collected byfiltration, washed with ether and dried under vacuum to give6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)-3,4-dihydroquinazolin-4-one(910 mg, 83%).

MS—ESI: 304 [MH]⁺

¹H NMR Spectrum: (DMSOd₆) 1.3-1.45 (m, 2H), 1.75 (d, 2H), 1.7-1.85 (m,1H), 1.9 (t, 2H), 2.2 (s, 3H), 2.8 (d, 2H), 3.9 (s, 3H), 4.0 (d, 2H),7.13 (s, 1H), 7.45 (s, 111), 7.99 (s, 1H)

EXAMPLE 3a

3.5M Hydrogen chloride in ethanol (75 μl, 0.26 mmol) was added to asuspension of4-chloro-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline (80 mg,0.25 mmol), (prepared as described for the starting material in Example2c), in isopropanol (3 ml), the mixture was heated to 50° C. and4-chloro-2-fluoroaniline (44 mg, 0.3 mmol) was added. The mixture washeated at reflux for 30 minutes. After cooling, the mixture was dilutedwith ether (3 ml). The precipitate was collected by filtration, washedwith ether and dried under vacuum to give4-(4-chloro-2-fluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazolinehydrochloride (105 mg, 82%).

MS—ESI: 431-433 [MH]⁺

The NMR spectrum of the protonated form of4-(4-chloro-2-fluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazolinehydrochloride shows the presence of two forms A and B in a ratio A:B ofapproximately 9:1.

¹H NMR Spectrum: (DMSO d₆; CF₃COOD) 1.55-1.7 (m, form A 2H), 1.85-2.0(m, form B 4H), 2.05 (d, form A 2H), 2.1-2.2 (m, form A 1H), 2.35 (s,3H); 2.79 (s, form A 3H), 2.82 (s, form B 3H), 3.03 (t, form A 2H),3.2-3.3 (m, form B 2H); 3.3-3.4 (m, form B 2H), 3.52 (d, form A 2H),4.02 (s, 3H), 4.13 (d, form A 2H), 4.3 (d, form B 2H), 7.41 (s, 1H),7.47 (dd, 1H), 7.63 (t, 1H), 7.69 (dd, 1H), 8.19 (s, 1H), 8.88 (s, 1H)

Elemental analysis: Found C 51.8 H 5.6 N 10.9 C₂₂H₂₄N₄O₂ClF 0.4H₂ORequires C 51.7 H 5.3 N 11.0% 2HCl

EXAMPLE 3b

An alternative method of preparation is as follows:

Triphenylphosphine (615 mg, 2.3 mmol) followed by diethylazodicarboxylate (3690, 2.3 mmol) were added to a solution of4-hydroxymethyl-l-methylpiperidine (151 mg, 1.1 mmol), (J Med. Chem1973, 16, 156), and4-(4-chloro-2-fluoroanilino)-7-hydroxy-6-methoxyquinazoline (250 mg,0.78 mmol), (prepared as described for the starting material in Example7), in methylene chloride (5 ml). After stirring for 30 minutes atambient temperature, 4-hydroxymethyl-1-methylpiperidine (51 mg, 0.39mmol), triphenylphosphine (102 mg, 0.39 mmol) and diethylazodicarboxylate (61 μl, 0.39 mmol) were added. After stirring for 15minutes, the volatiles were removed under vacuum and the residue waspurified by column chromatography eluting with methylenechloride/acetonitrile/methanol (70/10/20 followed by 75/5/20 and80/0/20). The fractions containing the expected product were combinedand the volatiles were removed by evaporation. The residue was dissolvedin a mixture of methylene chloride and methanol and 5M hydrogen chloridein isopropanol was added. The suspension was concentrated and the solidwas collected by filtration, washed with ether and dried under vacuum togive4-(4-chloro-2-fluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazolinehydrochloride (16 mg, 4%).

EXAMPLE 4

Under argon, sodium hydride (60%, 372 mg, 9.3 mmol) was added to asolution of 4-bromo-2,6-difluoroaniline (1.67 g, 8.08 mmol) in DMF.After stirring for 30 minutes at ambient temperature,4-chloro-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline (1.3 g,4.04 mmol) was added and stirring was continued for a further 20 hours.The mixture was poured onto water (130 ml) and extracted with ethylacetate. The organic layers were washed with water, brine, dried (MgSO₄)and the volatiles were removed by evaporation. The residue was purifiedby column chromatography on silica, eluting with methylenechloride/methanol (95/5) followed by methylene chloride/methanolcontaining ammonia (1%) (90/10). The fractions containing the expectedproduct were combined and evaporated. The residue was triturated withether, collected by filtration, washed with ether and dried under vacuumat 50° C. to give4-(4-bromo-2,6-difluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline(1.4 g, 70%).

MS—ESI: 493-495 [MH]⁺

¹H NMR Spectrum: (DMSOd₆) 1.3-1.45 (m, 2H), 1.8 (d, 2H), 1.7-1.9 (m,1H), 1.9 (t, 2H), 2.17 (s, 3H), 2.8 (d, 2H), 3.95 (s, 3H), 4.02 (d, 2H),7.2 (s, 1H), 7.63 (s, 1H), 7.6 (s, 1H), 7.82 (s, 1H), 8.35 (s, 1H)

Elemental analysis: Found C 53.8 H 4.8 N 11.3 C₂₂H₂₃N₄O₂BrF₂ Requires C53.6 H 4.7 N 11.4%

EXAMPLE 5

Using an analogous procedure to that described in Example 4,4-chloro-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline (246 mg,0.764 mmol), (prepared as described for the starting material in Example2c), was reacted with 4-chloro-2,6-difluoroaniline (250 mg, 1.53 mmol),(see WO 97/30035 Example 15), in DMF (9 ml) and in the presence ofsodium hydride (60%, 76.5 mg, 1.9 mmol) to give4-(4-chloro-2,6-difluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline(210 mg, 61%).

MS—ESI: 449-451 [MH]⁺

¹H NMR Spectrum: (DMSOd₆) 1.3-1.45 (m, 2H), 1.8 (d, 2H), 1.7-1.9 (m,1H), 1.9 (t, 2H), 2.2 (s, 3H), 2.8 (d, 2H), 3.96 (s, 3H), 4.02 (d, 2H),7.21 (s, 1H), 7.52 (s, 1H), 7.54 (s, 1H), 7.82 (s, 1H), 8.35 (s, 1H)

Elemental analysis: Found C 59.0 H 5.3 N 12.5 C₂₂H₂₃N₄O₂ClF₂ Requires C58.9 H 5.2 N 12.5%

The starting material was prepared as follows:

4-Chloro-2,6-difluoroaniline hydrochloride (see WO 97/30035 Example 15)was partitioned between methylene chloride and water and aqueous sodiumhydrogen carbonate was added until the pH of the aqueous layer wasapproximately 9. The organic layer was separated, washed with brine,dried (MgSO₄) and evaporated to give 4-chloro-2,6-difluoroaniline freebase.

EXAMPLE 6

A suspension of4-chloro-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline (200 mg,0.622 mmol), (prepared as described for the starting material in Example2c) and 2-fluoro-4-methylaniline (94 mg, 0.764 mmol) in isopropanol (5ml) containing 6.2M hydrogen chloride in isopropanol (110 μl) wasstirred at 80° C. for 1.5 hours. After cooling, the precipitate wascollected by filtration, washed with isopropanol, followed by ether anddried under vacuum. The solid was purified by column chromatography,eluting with methylene chloride/methanol (90/10) followed by 5% ammoniain methanol/methylene chloride (10/90). Evaporation of the fractionscontaining the expected product gave4-(2-fluoro-4-methylanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline(170 mg, 61%). MS—ESI: 411 [MH]⁺

¹H NMR Spectrum: (DMSOd₆) 1.3-1.45 (m, 2H), 1.8 (d, 2H), 1.7-1.9 (m,1H), 1.9 (t, 2H), 2.2 (s, 3H), 2.35 (s, 3H), 2.8 (d, 2H), 3.95 (s, 3H),4.01 (d, 2H), 7.1 (d, 1H), 7.13 (d, 1H), 7.16 (s, 1H), 7.4 (t, 1H), 7.81(s, 1H), 8.32 (s, 1H), 9.4 (s, 1H)

Elemental analysis: Found C 66.5 H 6.7 N 13.7 C₂₃H₂₇N₄O₂F 0.3H₂ORequires C 66.4 H 6.7 N 13.5%

EXAMPLE 7

1-tert-Butoxycarbonyl-4-hydroxymethylpiperidine (590 mg, 2.75 mmol),(prepared as described for the starting material in Example 1), followedby triphenylphosphine (1.2 g, 4.58 mmol) and diethyl azodicarboxylate(0.72 ml, 4.58 mmol) were added to a solution of4-(4-chloro-2-fluoroanilino)-7-hydroxy-6-methoxyquinazoline (585 mg,1.83 mmol) in methylene chloride (20 ml). After stirring for 1 hour atambient temperature, further triphenylphosphine (239 mg, 0.91 mmol) anddiethyl azodicarboxylate (0.14 ml, 0.91 mmol) were added. After stirringfor 1.5 hours, the volatiles were removed under vacuum and the residuewas purified by column chromatography eluting with ethylacetate/methylene chloride (1/1). The crude product was used directly inthe next step.

A solution of the crude product in methylene chloride (15 ml) containingTFA (4.5 ml) was stirred at ambient temperature for 1.5 hours. Thevolatiles were removed under vacuum. The residue was partitioned betweenwater and ethyl acetate. The aqueous layer was adjusted to pH9.5 with 2Nsodium hydroxide. The organic layer was separated, washed with water,followed by brine, dried (MgSO₄) and evaporated to give4-(4-chloro-2-fluoroanilino)-6-methoxy-7-(piperidin-4-ylmethoxy)quinazoline.

MS—ESI: 417-419 [MH]⁺

NMR Spectrum: (DMSOd₆) 1.1-1.3 (m, 2H), 1.75 (d, 2H), 1.85-2.0 (br s,1H), 2.55 (d, 2H), 2.95 (d, 2H), 3.95 (s, 3H), 4.0 (d, 2H), 7.2 (s, 1H),7.35 (dd, 1H), 7.55 (dd, 1H), 7.6 (t, 1H), 7.8 (s, 1H), 8.35 (s, 1H),9.55 (s, 1H)

The hydrochloride salt was made as follows:

4-(4-Chloro-2-fluoroanilino)-6-methoxy-7-(piperidin-4-ylmethoxy)quinazolinewas dissolved in a mixture of methanollmethylene chloride and 6Mhydrogen chloride in ether was added. The volatiles were removed undervacuum, the residue was triturated with ether, collected by filtration,washed with ether and dried under vacuum to give4-(4-chloro-2-fluoro)-6-methoxy-7-(piperidin-4-ylmethoxy)quinazolinehydrochloride (390 mg, 47% over 2 steps).

Elemental analysis: Found C 50.4 H 5.2 N 11.0 C₂₁H₂₂O₂N₄ClF 2.25HClRequires C 50.6 H 4.9 N 11.2%

The starting material was prepared as follows:

A solution of 7-benzyloxy-4-chloro-6-methoxyquinazoline hydrochloride(1.2 g, 4 mmol), (prepared as described in WO 97/22596, Example 1), and4-chloro-2-fluoroaniline (444 μl, 4 mmol) in 2-propanol (40 ml) washeated at reflux for 1.5 hours. After cooling, the precipitate wascollected by filtration, washed with 2-propanol then ether and driedunder vacuum to give7-benzyloxy-4-(4-chloro-2-fluoroanilino)-6-methoxyquinazolinehydrochloride (1.13 g, 64%).

m.p. 239-242° C.

¹H NMR Spectrum: (DMSOd₆) 4.0(s, 3H); 5.36(s, 2H); 7.39-7.52(m, 9H);8.1(s, 1H); 8.75(s, 1H)

MS—ESI: 410 [MH]⁺

Elemental analysis: Found C 59.2 H 4.3 N 9.4 C₂₂H₁₇N₃O₂ClF 1HCl RequiresC 59.2 H 4.1 N 9.4%

A solution of7-benzyloxy-4-(4-chloro-2-fluoroanilino)-6-methoxyquinazolinehydrochloride (892 mg, 2 mmol) in TFA (10 ml) was heated at reflux for50 minutes. After cooling, the mixture was poured onto ice. Theprecipitate was collected by filtration, dissolved in methanol (10 ml)and basified to pH11 with aqueous ammonia. After concentration byevaporation, the solid product was collected by filtration, washed withwater then ether and dried under vacuum to give4-(4-chloro-2-fluoroanilino)-7-hydroxy-6-methoxyquinazoline as a yellowsolid (460 mg, 72%).

m.p. 141-143° C.

MS—ESI: 320-322 [MH]⁺

¹H NMR Spectrum: (DMSOd₆) 3.95(s, 3H); 7.05(s, 1H); 7.35(d, 1H);7.54-7.59(m, 2H); 7.78(s, 1H); 8.29(s, 1H)

EXAMPLE 8

A suspension of7-(1-(tert-butoxycarbonyl)piperidin-4-ylmethoxy)-4-(2-fluoro-4-methylanilino)-6-methoxyquinazoline(318 mg, 0.64 mmol) in methylene chloride (5 ml) containing TFA (2.5 ml)was stirred at ambient temperature for 2 hours. The volatiles wereremoved under vacuum and the residue was partitioned between methylenechloride and water. The aqueous layer was adjusted to pH10-11. Theorganic layer was separated, washed with water, brine, dried (MgSO₄) andthe volatiles were removed by evaporation to give4-(2-fluoro-4-methylanilino)-6-methoxy-7-(piperidin-4-ylmethoxy)quinazoline(220 mg, 87%).

MS—ESI: 397 [MH]⁺

¹H NMR Spectrum: (DMSOd₆) 1.15-1.3 (m, 2H); 1.75 (d, 2H); 1.85-2.0 (m,1H); 2.4 (s, 3H); 3.0 (d, 2H); 3.3-3.4 (d, 2H); 3.95 (s, 3H); 4.0 (d,2H); 7.04 (d, 1H); 7.15 (d, 1H); 7.17 (s, 1H); 7.4 (t, 1H); 7.8 (s, 1H);8.3 (s, 1H); 9.4 (s, 1H)

The starting material was prepared as follows:

Using an analogous procedure to that described in Example 6,7-benzyloxy-4-chloro-6-methoxyquinazoline hydrochloride (1.55 g, 5.15mmol), (prepared, for example, as described in WO 97/22596, Example 1),was reacted with 2-fluoro-4-methylaniline (700 mg, 5.67 mmol) inisopropanol (90 ml) containing 6.2M hydrogen chloride in isopropanol (80μl, 0.51 mmol) to give7-benzyloxy-4-(2-fluoro-4-methylanilino)-6-methoxyquinazolinehydrochloride (2 g, 91%).

MS - ESI: 390 [MH]⁺

¹H NMR Spectrum: (DMSOd₆) 2.4 (s, 3H), 4.01 (s, 3H), 7.15 (d, 1H), 7.25(d, 1H), 7.35-7.6 (m, 7H), 8.3 (s, 1H), 8.78 (s, 1H)

A solution of7-benzyloxy-4-(2-fluoro-4-methylanilino)-6-methoxyquinazolinehydrochloride (2 g, 4.7 mmol) in TFA (20 ml) was heated at 80° C. for 5hours and stirred at ambient temperature overnight. The volatiles wereremoved under vacuum and the residue was suspended in water (50 ml).Solid sodium hydrogen carbonate was added until the pH was approximately7. The precipitate was then collected by filtration, washed with waterand dried under vacuum. The solid was purified by column chromatographyeluting with methanol/methylene chloride (5/95). After removal of thesolvent by evaporation, the solid was triturated with ether, collectedby filtration, washed with ether and dried under vacuum to give4-(2-fluoro-4-methylanilino)-7-hydroxy-6-methoxyquinazoline (1.04 g,74%).

MS—ESI: 300 [MH]⁺

¹H NMR Spectrum: (DMSOd₆) 2.4 (s, 3H), 4.0 (s, 3H), 7.15 (d, 1H), 7.22(s, 111), 7.25 (d, 1H), 7.41 (t, 1H), 8.05 (s, 1H), 8.7 (s, 1H), 11.0(s, 1H), 11.5-11.8 (br s, 1H)

Triphenylphosphine (2.19 g, 8.36 mmol) was added to a suspension of4-(2-fluoro-4-methylanilino)-7-hydroxy-6-methoxyquinazoline (1 g, 3.34mmol) in methylene chloride (10 ml) cooled at 0° C., followed by1-(tert-butoxycarbonyl)-4-hydroxymethylpiperidine (1.08 g, 5.01 mmol),(prepared as described for the starting material in Example 1), anddiethyl azodicarboxylate (1.31 ml, 8.36 mmol). After stirring for 2hours at ambient temperature, the volatiles were removed under vacuum.The residue was purified by column chromatography eluting with methylenechloride/methanol (2/98). After removal of the solvent by evaporation,the residue was triturated with ether, collected by filtration, washedwith ether and dried under vacuum to give7-(1-(tert-butoxycarbonyl)piperidin-4-ylmethoxy)-4-(2-fluoro-4-methylanilino)-6-methoxyquinazoline(327 mg, 20%).

MS - ESI: 497 [MH]⁺

¹H NMR Spectrum: (DMSOd₆) 1.15-1.3 (m, 2H); 1.45 (s, 9H); 1.8 (d, 2H);2.0-2.1 (m, 1H); 2.4 (s, 3H); 2.75-2.9 (br s, 2H); 3.95 (s, 3H); 4.0 (brs, 2H); 4.05 (d, 2H); 7.1 (d, 1H); 7.15 (d, 1H); 7.2 (s, 1H); 7.4 (t,1H); 7.85 (t, 1H); 8.32 (s, 1H); 9.45 (s, 1H)

EXAMPLE 9

A solution of4-(4-bromo-2,6-difluoroanilino)-7-(1-(tert-butoxycarbonyl)piperidin-4-ylmethoxy)-6-methoxyquinazoline(578 mg, lmmol) in methylene chloride (10 ml) containing TFA (4 ml) wasstirred at ambient temperature for 2 hours. The volatiles were removedunder vacuum and the residue was suspended in water. The aqueous layerwas adjusted to approximately pH10 and was extracted with methylenechloride. The organic layer was washed with water, brine , dried (MgSO₄)and the volatiles were removed by evaporation. The residue wastriturated with ether and dried under vacuum to give4-(4-bromo-2,6-difluoroanilino)-6-methoxy-7-(piperidin-4-ylmethoxy)quinazoline(110 mg, 23%).

MS—ESI: 479-481 [MH]+

¹H NMR Spectrum: (DMSOd₆) 1.15-1.3 (m, 2H); 1.75 (d, 2H); 1.85-2.0 (brs, 1H); 2.5 (d, 2H); 3.0 (d, 2H); 3.97 (s, 3H); 4.0 (d, 2H); 7.2 (s,1H); 7.62 (d, 2H); 7.82 (s, 1H); 8.35 (s, 1H)

¹H NMR Spectrum: (DMSOd₆; CF₃COOD) 1.5-1.65 (m, 2H); 2.0 (d, 2H);2.15-2.3 (br s, 1H); 3.0 (t, 2H); 3.4 (d, 2H); 4.02 (s, 3H); 4.15 (d,2H); 7.4 (s, 1H); 7.75 (d, 2H); 8.1 (s, 1H); 8.92 (s, 1H)

The starting material was prepared as follows:

Sodium hydride (60%, 612 mg, 15.3 mmol) was added to a solution of4-bromo-2,6-difluoroaniline (2.77 g, 6.65 mmol) in DMF (80 ml). Afterstirring for 30 minutes at ambient temperature,7-benzyloxy-4-chloro-6-methoxyquinazoline (2 g, 6.65 mmol), (prepared,for example, as described in WO 97/22596, Example 1, but the free basewas generated prior to 10 use), was added and stirring was maintainedfor 4 hours. The mixture was partitioned between ethyl acetate and water(200 ml). The organic layer was separated, washed with water, brine,dried (MgSO₄) and the volatiles were removed by evaporation. The residuewas triturated with isopropanol, collected by filtration, washed withether and dried under vacuum to give7-benzyloxy-4-(4-bromo-2,6-difluoroanilino)-6-methoxyquinazoline (1.95g, 62%).

MS—ESI: 472-474 [MH]⁺

¹H NMR Spectrum: (DMSOd₆) 3.94 (s, 3H), 5.3 (s, 2H), 7.3 (s, 1H), 7.4(d, 1H), 7.45 (t, 2H), 7.5 (s, 1H), 7.55 (d, 1H), 7.65 (d, 2H), 7.85 (s,1H), 8.35 (s, 1H), 9.4-9.6 (br s, 1H)

Using an analogous procedure to that described for the synthesis of thestarting material in Example 8,7-benzyloxy-4-(4-bromo-2,6-difluoroanilino)-6-methoxyquinazoline (1.9 g,4.02 mmol) was reacted with TFA (20 ml) to give4-(4-bromo-2,6-difluoroanilino)-7-hydroxy-6-methoxyquinazoline (1.5 g,98%).

¹H NMR Spectrum: (DMSOd₆) 3.95 (s, 3H), 7.1 (s, 1H), 7.6 (s, 1H), 7.65(s, 1H), 7.8 (s, 1H), 8.3 (s, 1H), 9.45 (br s, 1H), 10.5 (br s, 1H)

Using an analogous procedure to that described in the preparation of thestarting material in Example 8,4-(4-bromo-2,6-difluoroanilino)-7-hydroxy-6-methoxyquinazoline (1 g,2.62 mmol) was reacted with1-(tert-butoxycarbonyl)-4-hydroxymethylpiperidine (845 mg, 3.93 mmol),(prepared as described for the starting material in Example 1), to give4-(4-bromo-2,6-difluoroanilino)-7-(1-(tert-butoxycarbonyl)piperidin-4-ylmethoxy)-6-methoxyquinazoline(620 mg , 41%).

MS—ESI: 579-581 [MH]+

¹H NMR Spectrum: (DMSOd₆) 1.15-1.3 (m, 2H); 1.45 (s, 9H); 1.8 (d, 2H);2.0-2.1 (m, 1H); 2.7-2.9 (m, 2H); 3.95 (s, 3H); 4.0 (br s, 2H); 4.05 (d,2H); 7.22 (s, 1H); 7.65 (d, 2H); 7.85 (s, 1H); 8.35 (s, 1H); 9.4-9.6 (brs, 1H)

EXAMPLE 10

Using an analogous procedure to that described in Example 9,7-(1-(tert-butoxycarbonyl)piperidin-4-ylmethoxy)-4-(4-chloro-2,6-difluoroanilino)-6-methoxyquinazoline(95 mg, 0.2 mmol) in methylene chloride (2 ml) was treated with TFA (800μl) to give4-(4-chloro-2,6-difluoroanilino)-6-methoxy-7-(piperidin-4-ylmethoxy)quinazoline(20mg, 26%).

MS—ESI: 435-437 [MH]+

¹H NMR Spectrum: (DMSOd₆) 1.2-1.3 (m, 2H); 1.75 (d, 2H); 1.85-2.0 (br s,1H); 2.5 (d, 2H); 3.0 (d, 2H); 3.97 (s, 3H); 4.0 (d, 2H); 7.2 (s, 1H);752 (d, 2H); 7.85 (s, 1H); 8.35 (s, 1H)

The starting material was prepared as follows:

Using an analogous procedure to that described for the preparation ofthe starting material in Example 9,7-benzyloxy-4-chloro-6-methoxyquinazoline (184 mg, 0.61 mmol),(prepared, for example, as described in WO 97/22596, Example 1, but thefree base was generated prior to use), was reacted with4-chloro-2,6-difluoroaniline (200 mg, 1.22 mmol) in the presence ofsodium hydride (60%, 87 mg, 1.4 mmol) in DMF (8 ml) to give7-benzyloxy-4-(4-chloro-2,6-difluoroanilino)-6-methoxyquinazoline (212mg, 74%).

MS—ESI: 428 [MH]+

¹H NMR Spectrum: (DMSOd₆) 3.96 (s, 3H); 5.31 (s, 2H); 7.32 (s, 1H); 7.4(d, 1H); 7.45 (t, 2H); 7.5-7.6 (m, 4H); 7.85 (s, 1H); 8.35 (br s, 1H);9.55 (br s, 1H)

A solution of7-benzyloxy-4-(4-chloro-2,6-difluoroanilino)-6-methoxyquinazoline (200mg, 0.47 mmol) in TFA (3 ml) was stirred at 80° C. for 3 hours. Aftercooling, the volatiles were removed under vacuum and the residue wasdissolved in water containing 5% methanol. The pH was adjusted to 8 withsodium hydrogen carbonate and the solid was collected by filtration andwashed with water. The solid was solubilised in a mixture of ethylacetate/methanol/methylene chloride (47/6/47). The volatiles wereremoved under vacuum to give4-(4-chloro-2,6-difluoroanilino)-7-hydroxy-6-methoxyquinazoline (126 mg,80%).

MS - ESI: 338 [MH]+

¹H NMR Spectrum: (DMSO d₆) 3.95 (s, 3H); 7.1 (s, 1H); 7.55 (d, 2H); 7.8(s, 1H); 8.3 (s, 1H); 9.42 (br s, 1H)

Using an analogous procedure to that described for the preparation ofthe starting material in Example 9,4-(4-chloro-2,6-difluoroanilino)-7-hydroxy-6-methoxyquinazoline (150 mg,0.44 mmol) was reacted with1-(tent-butoxycarbonyl)-4-hydroxymethylpiperidine (150 mg, 0.88 mmol),(prepared as described for the starting material in Example 1), to give7-(1-(tert-butoxycarbonyl)piperidin-4-ylmethoxy)-4-(4-chloro-2,6-difluoroanilino)-6-methoxyquinazoline(113 mg, 59%).

MS - ESI: 535 [MH]+

¹H NMR Spectrum: (DMSOd₆) 1.15-1.3 (m, 2H); 1.45 (s, 9H); 1.8 (d, 2H);2.0-2.1 (m, 1H); 2.7-2.9 (m, 2H); 3.95 (s, 3H); 4.0 (br s, 2H); 4.05 (d,2H); 7.2 (s, 1H); 7.6 (m, 2H); 7.8 (s, 1H); 8.35 (s, 1H); 9.4-9.6 (br s,1H)

EXAMPLE 11

The following illustrate representative pharmaceutical dosage formscontaining the compound of formula I, or a pharmaceutically acceptablesalt thereof (hereafter compound X), for therapeutic or prophylactic usein humans:

(a) Tablet I mg/tablet Compound X 100 Lactose Ph. Eur 182.75Croscarmellose sodium 12.0 Maize starch paste (5% w/v paste) 2.25Magnesium stearate 3.0 (b) Tablet II mg/tablet Compound X 50 Lactose Ph.Eur 223.75 Croscarmellose sodium 6.0 Maize starch 15.0Polyvinylpyrrolidone (5% w/v paste) 2.25 Magnesium stearate 3.0 (c)Tablet III mg/tablet Compound X 1.0 Lactose Ph. Eur 93.25 Croscarmellosesodium 4.0 Maize starch paste (5% w/v paste) 0.75 Magnesium stearate 1.0(d) Capsule mg/capsule Compound X 10 Lactose Ph. Eur 488.5 Magnesiumstearate 1.5 (e) Injection I (50 mg/ml) Compound X 5.0% w/v 1M Sodiumhydroxide solution 15.0% v/v  0.1M Hydrochloric acid (to adjust pH to7.6) Polyethylene glycol 400 4.5% w/v Water for injection to 100% (f)Injection II 10 mg/ml) Compound X 1.0% w/v Sodium phosphate BP 3.6% w/v0.1M Sodium hydroxide solution 15.0% v/v  Water for injection to 100%(g) Injection III (1 mg/ml, buffered to pH6) Compound X 0.1% w/v Sodiumphosphate BP 2.26% w/v  Citric acid 0.38% w/v  Polyethylene glycol 4003.5% w/v Water for injection to 100% Note The above formulations may beobtained by conventional procedures well known in the pharmaceuticalart. The tablets (a)-(c) may be enteric coated by conventional means,for example to provide a coating of cellulose acetate phthalate.

1. A quinazoline derivative of the formula I:

wherein: m is an integer from 1 to 3; R¹ represents halogeno orC₁₋₃alkyl; X¹ represents —O—; R² is selected from one of the followingthree groups: 1) C₁₋₅alkylR³ (wherein R³ is piperidin-4-yl which maybear one or two substituents selected from hydroxy, halogeno, C₁₋₄alkyl,C₁₋₄hydroxyalkyl and C₁₋₄alkoxy; 2) C₂₋₅alkenylR³ (wherein R³ is asdefined herein); 3) C₂₋₅alkynylR³ (wherein R³ is as defined herein); andwherein any alkyl, alkenyl or alkynyl group may bear one or moresubstituents selected from hydroxy, halogeno and amino; or a saltthereof.
 2. A quinazoline derivative as claimed in claim 1 wherein thephenyl group bearing (R¹)_(m) is selected from 2-fluoro-4-methylphenyl,4-chloro-2,6-difluorophenyl, 4-bromo-2,6-difluorophenyl,4-chloro-2-fluorophenyl group and 4-bromo-2-fluorophenyl.
 3. Aquinazoline derivative as claimed in claim 1 or claim 2 wherein R² isC₁₋₅alkylR³ (wherein R³ is as defined in claim 1).
 4. A quinazolinederivative as claimed in claim 1 wherein R² is piperidin-4-ylmethyl inwhich the piperidine ring may bear one or two substituents selected fromC,4alkyl.
 5. A quinazoline derivative as claimed in claim 1 of theformula II:

wherein: ma is an integer from 1 to 3; R^(1a) represents halogeno orC₁₋₃alkyl; X^(1a) represents —O—; R^(2a) is selected from one of thefollowing three groups: 1) C₁₋₃alkylR³ (wherein R³ is as defined inclaim 1); 2) C₂₋₅alkenylR³ (wherein R³ is as defined in claim 1); 3)C₂₋₅alkynylR³ (wherein R³ is as defined in claim 1); or a salt thereof.6. A quinazoline derivative as claimed in claim 1 selected from:4-(4-chloro-2-fluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline,4-(2-fluoro-4-methylanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline,4-(4-bromo-2-fluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline,4-(4-chloro-2,6-difluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline,4-(4-bromo-2,6-difluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline,4-(4-chloro-2-fluoroanilino)-6-methoxy-7-(piperidin-4-ylmethoxy)quinazoline,4-(2-fluoro-4-methylanilino)-6-methoxy-7-(piperidin-4-ylmethoxy)quinazoline,4-(4-bromo-2-fluoroanilino)-6-methoxy-7-(piperidin-4-ylmethoxy)quinazoline,4-(4-chloro-2,6-difluoroanilino)-6-methoxy-7-(piperidin-4-ylmethoxy)quinazoline,and4-(4-bromo-2,6-difluoroanilino)-6-methoxy-7-(piperidin-4-ylmethoxy)quinazoline,and salts thereof.
 7. A quinazoline derivative as ed in claim 1 selectedfrom:4-(4-chloro-2-fluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline,4-(4-bromo-2-fluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline,4-(4-chloro-2, 6-difluoroanilino)-6-methoxy-7-(1 -methyIpiperidin-4-ylmethoxy)quinazoline, and4-(4-bromo-2,6-difluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline, and salts thereof.
 8. Aquinazoline derivative as claimed in claim 1 selected from:4-(4-bromo-2-fluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazolineand salts thereof.
 9. A quinazoline derivative as claimed in claim 1 inthe form of a pharmaceutically acceptable salt.
 10. A process for thepreparation of a quinazoline derivative of formula I, as defined inclaim 1, or a salt thereof which comprises: (a) the reaction of acompound of the formula III:

wherein R² and X¹ are as defined in claims 1 and L¹ is a displaceablemoiety, with a compound of the formula IV:

wherein R¹ and m are as defined in claim 1; (b) the reaction of acompound of the formula V:

wherein m, X¹ and R are as defined in claim 1 with a compound of formulaVI:R²-L¹   (VI) wherein R² is as defined in claims 1 and L¹ is as definedherein; (c) the reaction of a compound of the formula VII:

with a compound of the formula VIII:R²—X¹—H   (VIII) wherein R¹, R², m and X¹ are as defined in claim 1 andL¹ is as defined herein; or (d) the deprotection of a compound of theformula IX:

wherein R¹, m and X¹ are as defined in claim 1, and R⁴ represents aprotected R² group wherein R² is as defined in claim 1 but additionallybears one or more protecting groups P²; and when a pharmaceuticallyacceptable salt of a quinazoline derivative of formula I is required,reaction of the compound obtained with an acid or base whereby to obtainthe desired pharmaceutically acceptable salt.
 11. A pharmaceuticalcomposition which comprises as active ingredient a compound of formula Ias defined in claim 1 or a pharmaceutically acceptable salt thereof, inassociation with a pharmaceutically acceptable excipient or carrier. 12.(canceled)
 13. A method for producing an antiangiogenic and/or vascularpermeability reducing effect in a warm-blooded animal in need of suchtreatment which comprises administering to said animal an effectiveamount of a compound of formula I as defined in claim 1 or apharmaceutically acceptable salt thereof.